Toksik Antarif Fix

Tugas Toksikologi dan Kesehatan Lingkungan
MEKANISME TERJADINYA KANKER OLEH PCBS
Dosen:
Dr. Ir. Etty Riani, MS
Oleh :
Antarif Kusuma Brata
P052150341
PROGRAM STUDI MAGISTER

PENGELOLAAN SUMBERDAYA ALAM DAN LINGKUNGAN
SEKOLAH PASCASARJANA
INSTITUT PERTANIAN BOGOR
2016
1. PENDAHULUAN
1.1. Latar Belakang
Bahan Berbahaya dan Beracun (B3) merupakan bahan yang karena sifat atau
konsentrasi, jumlahnya, baik secara langsung maupun tidak langsung, dapat
mencemari atau merusak lingkungan hidup, kesehatan, kelangsungan hidup manusia
serta mahluk hidup lain. Menurut data dari Environmental Protection Agency (EPA)
tahun 1997, yang menyusun top-20 B3 antara lain: Arsenic, Lead, Mercury, Vinyl
chloride, Benzene, Polychlorinated Biphenyls (PCBs), Kadmium, Benzo(a)pyrene,
Benzo(b)fluoranthene, Polycyclic Aromatic Hydrocarbons, Chloroform, Aroclor
1254,
DDT,
Aroclor
Dibenz[a,h]anthracene,
1260,
Dieldrin,
Trichloroethylene,
Chromium
Hexachlorobutadiene,
(hexa
Chlordane.
valent),
Beberapa
diantaranya merupakan logam berat, antara lain Arsenic (As), Lead (Pb), Mercury
(Hg), Kadmium (Cd) dan Chromium (Cr) (Sudarmaji, 2006). Logam-logam berat
tersebut dalam konsentrasi tinggi akan berbahaya bagi kesehatan manusia bila
ditemukan di dalam lingkungan, baik di dalam air, tanah maupun udara.
Polychlorinated biphenyls (PCBs) adalah suatu substansi kimia organik sintetis
yang dikenal sebagai hydrocarbon chlorinated. PCBs bersifat persisten jika dilepaskan
ke lingkungan karena ketahanannya terhadap proses metabolisme yang dapat
memecahkan mereka ke bentuk komposisi kimia yang lebih sederhana. Solubilitasnya
yang rendah di air menyebabkan PCBs terkamulasi dalam jaringan lemak manusia dan
hewan. PCBs dikenal menyebabkan efek kronik pada organ reproduksi, kekacauan
pencernaan, dan luka pada hewan laboratorium/percobaan. Sebagai tambahan EPA
mencurigai PCBs sebagai karsinogen pada manusia. Ditemukan dalam beberapa
penelitian bahwa senyawa PCBs memiliki sifat aktif memicu pembentukan dan
perkembangan sel kanker.
Makalah ini akan membahas lebih lanjut mengenai senyawa PCBs itu sendiri,
mulai dari pengertian, karakteristik, dan juga bagaimana mekanisme PCBs di dalam
tubuh sebagai zat bersifat karsinogenik sehingga menyebabkan kanker.
1.2. Rumusan Masalah

Sebelum menentukan tujuan yang ingin dicapai dalam penulisan makalah ini perlu
adanya perumusan masalah guna menentukan fokus bahasan. Masalah dalam tulisan ini
adalah pentingnya mengetahui apakah Kanker itu, senyawa PCBs dan juga
mekanismenya dalam menyebabkan terjadinya kanker dalam tubuh manusia
1.3.Tujuan
Secara umum tujuan dari makalah ini adalah pemaparan mengenai kanker dan
pembentukannya,
serta
profil
senyawa
PCBs
dan
sifat
mekanisme
sifat
karsinogenesisnya.
1.4.Metoda
Metode yang digunakan dalam penulisan ini adalah studi literatur serta telaah dari
berbagai sumber literatur yang digunakan
2. PEMBAHASAN
2.1.
Kanker dan Karsinogenesis

Menurut WHO, kanker adalah istilah umum untuk satu kelompok besar penyakit
yang dapat mempengaruhi setiap bagian dari tubuh. Istilah lain yang digunakan adalah
tumor ganas dan neoplasma. Salah satu fitur mendefinisikan kanker adalah pertumbuhan
sel-sel baru secara abnormal yang tumbuh melampaui batas normal, dan yang kemudian
dapat menyerang bagian sebelah tubuh dan menyebar ke organ lain. Proses ini disebut
metastasis. Metastasis merupakan penyebab utama kematian akibat kanker (WHO 2009).
Menurut National Cancer Institute (2009), kanker adalah suatu istilah untuk penyakit di
mana sel-sel membelah secara abnormal tanpa kontrol dan dapat menyerang jaringan di
sekitarnya. Kanker adalah istilah umum yang dipakai untuk menunjukkan neoplasma
ganas, dan ada banyak tumor atau neoplasma lain yang tidak bersifat kanker (Price et al.,
2006). Neoplasma secara harfiah berarti pertumbuhan baru. Suatu neoplasma, sesuai
definisi Wills, adalah massa abnormal jaringan yang pertumbuhannya berlebihan dan
tidak terkoordinasikan dengan pertumbuhan jaringan normal serta terus demikian
walaupun rangsangan yang memicu perubahan tersebut telah berhenti (Kumar et al.,
2007).
Pada umumnya, kanker timbul karena paparan terhadap suatu karsinogen secara
berkali-kali dan aditif pada dosis tertentu, tetapi pada keadaan tertentu dapat juga timbul
dari dosis tunggal karsinogen. Penyebab kanker dapat satu karsinogen yang sama
misalnya asap rokok (kanker paru), dapat dua karsinogen yang berlainan misalnya asap
rokok dan debu asbes (kanker paru), asap rokok dan radiasi sinar X (kanker paru), asap
rokok dan alkohol (kanker orofarings, larings dan esofagus), gen kanker dan karsinogen
lingkungan.
Beberapa macam kanker terjadi dari satu faktor yang dominan misalnya sinar
ultraviolet yang menimbulkan kanker kulit dan kelainan kromosom yang menimbulkan
retinoblastoma. Karsinogenesis yang diinduksi karsinogen kimia atau fisik maupun
biologik memerlukan waktu yang disebut periode laten yaitu waktu dari pertama kali
terpapar suatu karsinogen sampai terlihat kanker secara klinis.
Periode laten dari
kebanyakan kanker seringkali 20 tahun atau lebih. Efek karsinogen yang lemah dapat
tidak terlihat, sebab periode latennya melampaui masa hidup seseorang. Karsinogenesis
dapat dibagi dalam tiga fase utama yaitu fase inisiasi, promosi dan progresi (Kartawiguna
2001).
a. Fase inisiasi
Fase ini berlangsung cepat. Karsinogen kimia misalnya golongan alkylating dapat
langsung menyerang tempat dalam molekul yang banyak elektronnya, disebut karsinogen
nukleofilik. Karsinogen golongan lain misalnya golongan polycyclic aromatic
hydrocarbon sebelum menyerang dikonversikan (diaktifkan) dulu secara metabolik
(kimiawi) menjadi bentuk defisit elektron yang disebut karsinogen elektrofilik reaktif.
Tempat yang diserang adalah asam nukleat (DNA/ RNA) atau protein dalam sel terutama
di atom nitrogen, oksigen dan sulfur. Air dan glutation juga diserang, dalam beberapa
kasus reaksi ini di-katalisasi oleh enzim seperti glutathione-S-transferase. Ikatan
karsinogen dengan DNA menghasilkan lesi di materi genetik. RNA yang berikatan dengan
karsinogen bermodifikasi menjadi DNA yang dimutasi. Karsinogen kimia yang berikatan
dengan DNA disebut genotoksik dan yang tidak berikatan dengan DNA disebut epigenetik.
Karsinogen genotoksik dapat juga mempunyai efek epigenetik. Kokarsinogen dan
promotor termasuk dalam karsinogen epigenetik yang menyebabkan kerusakan jaringan
kronis, perubahan sistem imun tubuh, perubahan hormon atau berikatan dengan protein
yang represif terhadap gen tertentu. Jadi karsinogen epigenetik dapat mengubah kondisi
lingkungan sehingga fungsi sebuah gen berubah, bukan strukturnya. Waktu yang
dibutuhkan dari pertama kali sel diserang karsinogen sampai terbentuk lesi di materi
genetik adalah beberapa menit. Sel berusaha mengoreksi lesi ini dengan detoksifikasi
kemudian diekskresi atau dapat terjadi kematian sel atau terjadi reparasi DNA yang rusak
tersebut oleh enzim sel menjadi sel normal kembali. Karsinogen kimia dapat
didetoksifikasi/ dinon-aktifkan kemudian diekskresi atau dapat langsung diekskresi.
Tetapi dari proses pengnon-aktifan ini dapat terbentuk metabolit yang karsinogenik.
Sebelum terjadi reparasi DNA dapat terjadi replikasi DNA yaitu satu siklus proliferasi sel
yang menyebabkan lesi DNA tersebut menjadi permanen disebut fiksasi lesi. Waktu yang
dibutuhkan dari pertama kali sel diserang karsinogen sampai terjadi fiksasi lesi (terbentuk
sel terinisiasi) adalah beberapa hari (1-2 hari). Replikasi DNA terjadi karena terdapatnya
sel nekrotik sebagai akibat karsinogen. Replikasi ini dapat diinduksi oleh lain bahan kimia
toksik, bakteri (misalnya colitis ulcerativa menjadi kanker kolon, bronkitis kronis menjadi
kanker paru pada perokok), virus, parasit (schistosomiasis di Afrika menjadi kanker
kandung kemih), defisiensi diet tertentu, hormon dan prosedur percobaan seperti
hepatektomi parsial. Pada jaringan yang mengalami peradangan atau sedang berproliferasi
(misalnya luka yang menyembuh) atau jaringan yang berproliferasi terus menerus
(misalnya sumsum tulang, epitel saluran pencernaan) tanpa terangsang dari luarpun dapat
terjadi replikasi DNA. Pada peradangan belum diketahui apakah terjadi akibat peradangan
membantu pertumbuhan sel atau melemahnya daya tahan tubuh. Sel terinisiasi dapat
mengalami kematian, bila tidak, maka sel dapat masuk ke fase promosi. Pada akhir fase
inisiasi belum terlihat perubahan histologis dan biokimiawi hanya terlihat nekrosis sel
dengan meningkatnya proliferasi sel.
b. Fase promosi
Sel terinisiasi dapat tetap tenang bila tidak dihidupkan oleh zat yang disebut
promotor. Promotor sendiri tidak dapat menginduksi perubahan kearah neoplasma
sebelum bekerja pada sel terinisiasi, hal ini telah dibuktikan pada percobaan binatang. Bila
promotor ditambahkan pada sel terinisiasi dalam kultur jaringan, sel ini akan
berproliferasi. Jadi promotor adalah zat proliferatif. Promosi adalah proses yang
menyebabkan sel terinisiasi berkembang menjadi sel preneoplasma oleh stimulus zat lain
(promotor). Pada percobaan binatang dibuktikan terdapat karsinogen kimia yang bekerja
sendiri sebagai inisiator dan promotor disebut karsinogen komplit. Dari penyelidikan pada
kultur jaringan diketahui fase ini berlangsung bertahun-tahun (10 tahun atau lebih) dan
reversibel sebelum terbentuknya sel tumor yang otonom.
c. Fase progresi
Fase ini berlangsung berbulan-bulan. Pada awal fase ini, sel preneoplasma dalam
stadium metaplasia berkembang progresif menjadi stadium displasia sebelum menjadi
neoplasma. Terjadi ekspansi populasi selsel ini secara spontan dan ireversibel. Sel-sel
menjadi kurang responsif terhadap sistem imunitas tubuh dan regulasi sel. Pada esofagus
epitel berlapis gepeng berubah atau metaplasia menjadi epitel selapis torak yang kemudian
berkembang menjadi jaringan dalam keadaan displasia yang kemudian berkembang
menjadi neoplasma. Pada kolon, polip adalah bentuk metaplasia. Pada tingkat metaplasia
dan permulaan displasia (ringan sampai sedang) masih bisa terjadi regresi atau remisi yang
spontan ke tingkat lebih awal yang frekwensinya semakin menurun dengan bertambahnya
progresivitas lesi tersebut. Belum banyak diketahui perubahan yang terjadi dan faktorfaktor yang mempengaruhinya. Batas yang pasti perubahan lesi preneoplasma menjadi
neoplasma sulit ditentukan. Pada akhir fase ini gambaran histologis dan klinis
menunjukkan keganasan. Penyelidikan terakhir memperlihatkan terjadi aglutinasi pada
permukaan sel kanker sehingga sel kanker tumbuh terus meskipun terjadi kontak antar sel.
Permukaan sel kanker mempunyai lebih sedikit neksus (daerah kontak antar sel). Ini
menunjukkan kurangnya metabolisme dan pertukaran ion-ion antar sel yang juga
menyebabkan sel kanker bertambah otonom. Hal ini lebih nyata pada keadaan displasia
yang progresif ke arah neoplasma. Semua perubahan struktur, metabolik dan kelakuan sel
ini terjadi karena mutasi yang mengenai inti, mitokondria dan membran endoplasma sel.
Kebanyakan sel kanker mensekresi enzim fibrinolitik yang melarutkan jaringan ikat di
sekitarnya dan faktor angiogenesis yang menginduksi pembentukan kapilar darah baru di
antara pembuluh darah yang berdekatan dengan sel kanker untuk nutrisinya. Pada
permukaan sel kanker terbentuk antigen yang menimbulkan respons imun selular dan
humoral untuk melawan sel kanker. Antigen permukaan ini sering ditemukan di jaringan
fetus, mempunyai hubungan dengan derajat diferensiasi sel dan kekhasannya dipakai
sebagai tambahan pada diagnostik kanker.
2.2. Polychlorinated biphenyls (PCBs)
Polychlorinated biphenyls (PCBs) adalah suatu substansi kimia organik sintetis
yang dikenal sebagai hydrocarbon chlorinated. PCBs bersifat persisten jika dilepaskan ke
lingkungan karena ketahanannya terhadap proses metabolisme yang dapat memecahkan
mereka ke bentuk komposisi kimia yang lebih sederhana. Solubilitasnya yang rendah di
air menyebabkan PCBs terkamulasi dalam jaringan lemak manusia dan hewan. PCBs
dikenal menyebabkan efek kronik pada organ reproduksi, kekacauan pencernaan, dan luka
pada hewan laboratorium/percobaan. Sebagai tambahan EPA mencurigai PCBs bersifat

karsinogen pada manusia. Struktur kimia Polychlorinated Biphenyls. Rumus kimia dari
senyawa PCBs adalah C12H10-nCl, dimana n memiliki kisaran 1 10, sehingga secara teori
terdapat kemungkinan 209 tipe PCBs. Semua tipe tersebut mempunyai struktur dasar
kimia yang sama seperti di bawah ini.
Gambar 1. Struktur dasar kimia Polychlorinated Biphenyls

PCBs mulai diperkenalkan dan diproduksi secara komersial sejak tahun 1920-an
untuk digunakan sebagai dielektrik pada transformer dan kapasitor, sebagai cairan
pendingin pada sistem hidraulik, pembuatan pelumas, bahan pembuatan pestisida, bahan
pemadam api, bahan campuran dalam cat, lem, penyalin dalam kertas, dan plastik
(Robertson & Ludewig 2011). Senyawa ini memiliki stabilitas yang baik, hal ini yang
membuat PCBs menjadi senyawa yang sangat diunggulkan dan memiliki pasar tersebar
di seluruh dunia. Produksi senyawa PCBs mencapai puncaknya pada tahun 1970-an,
namun mengalami penurunan setelah itu akibat mulai muncul pelarangan penggunaan
senyawa ini oleh beberapa negara karena telah mulai muncul penelitian dan penelitian
mengenai bahaya senyawa ini. Meskipun demikian senyawa ini masih tetap digunakan
hingga sekarang dan menjadi potensi bahaya bagi kesehatan lingkungan maupun manusia
itu sendiri (Hu & Hornbuckle 2010).
Tabel 1. IUPAC number dan posisi atom cl dalam segala jenis PCBs
Sifat senyawa PCBs yang stabil dipandang sebagai kelebihan dalam

pemanfaatannya di awal pemanfaatannya, namun belakangan sifat ini ternyata menjadi
sebuah ancaman sendiri seiring dengan peningkatan jumlah PCBs dimanfaatkan dan
terbuang ke lingkungan, senyawa ini menjadi senyawa yang membebani lingkungan
akibat tidak mudah terurai dan sifatnya sebagai senyawa terakumulasi. Akibatnya
senyawa ini terakumulasi di hampir semua badan lingkungan, baik itu air, udara maupun
tanah (Rodziewicz et al. 2004). PCBs belakangan juga ditemukan terakumulasi dalam
makanan, seperti ikan bahkan juga ditemukan dalam air susu ibu dan jaringan adiposa
manusia (Zhao et al. 2010). PCB sangat mudah menyebar, penyebaran paling banyak
didapatkan dari udara, air, bahkan dapat ditemukan akumulasi dalam bahan makanan yang
akan dikonsumsi oleh manusia.
Gambar 2. Siklus penyebaran PCBs di lingkungan
2.3.
Mekanisme Karsinogenisitas PCBs

PCBs telah dikategorikan oleh International Agency for Research on Cancer
(IARC) sebagai senyawa potensial penyebab kanker (IARC 1987). Di dalam studi yang
dilakukan IARC terdapat dua belas kasus yang menunjukan hubungan antara PCBs
dengan kanker. Salah satu contoh yang menunjukan adanya signifikansi nyata antara
PCBs dan kanker terdapat pada kasus kanker testikel di departemen keamanan U.S. yang
penelitiannya dilakukan oleh McGlynn et al. (2009). Terdapat beberapa kemungkinan
mekanisme PCB sebagai karsinogen menyebabkan kanker pada tubuh manusia,,
diantaranya:
Aktivasi metabolik dari PCBs menjadi senyawa reaktif mutagen penyebab
perubahan karyotipe (Robertson & Ludewig 2011)
Telah lama diketahui bahwa senyawa biphenyl dan biphenyl
terhalogenasi merupakan Hidroxylated in vivo dan in vitro. Reaksi hidroksilasi
ini umumnya terkatalisis oleh isoform cytochrome P-450. Eksperimen yang
pernah dilakukan menunjukan bahwa metabolisme dari PCB3 menghasilkan
adisi nukleotida in vitro dengan kecenderungan formasi purin dibanding
pyrimidin. Kebanyakan adisi ini berasal dari arene oxide intermediate. Hasil ini
menunjukan bahwa terdapat pengaruh PCB terhadap perubahan DNA melalui
proses metabolik aktivasi pembentukan senyawa intermediate yang bersifat
mutagen.
Secara garis besar gambaran proses mengenai mekanisme senyawa PCBs
dan senyawa kimia lain menjadi penanggung jawab terjadinya pembentukan sel
dapat dilihat pada Gambar 3.
Gambar 3. Proses mekanisme terjadinya kanker akibat PCB dan

senyawa Kimia Lain (Kira & Toren 2014)
Kerusakan Oksidatif DNA (Robertson & Ludewig 2011)

PCBs dengan kandungan klorin rendah menghasilkan Reactive Oxygen
Species (ROS) dan oxydative stress intraselular. Senyawa ini dikenal sebagai
radikal bebas yang sangat mudah mengganggu susunan jaringan hidup, terutama
hydroxyl radical yang dapat menyebabkan terjadinya pembentukan 8oxodeoxyguanosine (8-oxodG) yaitu ikatan DNA yang mutagenik menyebabkan
terjadinya perubahan struktur G menjadi T. Hydroxyl radical juga dapat
menyerang asam lemak (linoleic, linolinic, oleic, dan lainnya) dan membuat
pembentukan turunan lemak peroksida seperti acrolein, crotonaldehyde, dan
malondiadehyde. Ketiga lemak peroksida ini dapat merubah struktur DNA
sehingga menjadi DNA mutagenik yang akhirnya gagal dalam menyimpan dan
memberikan perintah kerja terhadap sel. Seperti diketahui, bila terjadi seperti itu,
kemungkinan sel akan tumbuh dan berkembang tidak secara semestinya dan
menjadi sel kanker.
Gambar 4. Mekanisme Kerusakan Oksidatif DNA (Kira & Toren 2014)
Senyawa PCBs sebagai senyawa karsinogen yang memiliki peran dalam pembentukan
kanker di dalam tubuh berperan dalam proses inisisasi dan promosi. Dengan dua
mekanisme utama di atas, adanya akumulasi PCBs dalam tubuh memperbesar
kemungkinan terjadinya mutasi DNA yang mengakibatkan kelainan dalam pembentukan
dan perkembangan sel, sehingga terbentuk jaringan abnormal yang berikutnya menjadi
kanker. Semakin besar jumlah akumulasi senyawa PCBs di dalam tubuh, semakin besar
juga kemungkinan terjadinya proses pembentukan sel mutasi penyebab kanker.
3. PENUTUP
Senyawa PCBs merupakan senyawa yang banyak digunakan dalam berbagai
kegiatan aktivitas manusia, mengingat sifatnya yang relatif sangat stabil, paling banyak
ditemukan dalam penggunaan sebagai pendingin trafo dan alat mesin lainnya. Namun, di
lain sisi senyawa ini juga memiliki ancaman tersendiri, karena sifatnya yang stabil
tersebut, senyawa ini tidak mudah hilang, sehingga keberadaannya terakumulasi di semua
bidang lingkungan. Di tubuh manusia sendiri juga terjadi akumulasi senyawa PCBs. jalur
masuk paling banyak senyawa PCBs masuk ke dalam tubuh manusia adalah melalui
akumulasi di bahan makanan. Selain itu jalur masuk PCBs ke dalam tubuh manusia dapat
berasal dari air dan udara. PCBs dalam beberapa penelitian terbukti memiliki sifat
karsinogen. Terdapat 2 mekanisme utama PCBs dalam karsinogenesis pembentukan
kanker, yaitu melalui aktivasi menjadi senyawa intermediates dan juga pembentukan
radikal bebas seperti ROS dan hydroxyl. PCBs dalam pembentukan sel kanker memiliki
peran dalam tahap inisiasi dan promosi sel kanker dalam tubuh.
DAFTAR PUSTAKA
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Zhao HX, AD Adamcakova, D Hu, KC Hornbuckle, CL Just, LW Robertson, PS Thorne,
HJ Lehmler. Development of synthetic PCB mixture resembling the average
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NIH Public Access

Author Manuscript
Environ Int. Author manuscript; available in PMC 2011 November 1.
NIH-PA Author Manuscript
Published in final edited form as:

Environ Int. 2010 November ; 36(8): 819827. doi:10.1016/j.envint.2009.03.003.
Development of a Synthetic PCB Mixture Resembling the Average

Polychlorinated Biphenyl Profile in Chicago Air
H. Zhao1, A. Adamcakova-Dodd1, D. Hu2, K.C. Hornbuckle2, C.L. Just2, L.W. Robertson1,
P.S. Thorne1, and H.-J. Lehmler1,*
1The University of Iowa, Department of Occupational and Environmental Health, 100 Oakdale
Campus, 124 IREH, Iowa City, IA, 52242, USA
2Department
of Civil and Environmental Engineering, University of Iowa, Iowa City, IA, 52242, USA
Abstract
Studies of environmental and toxic effects of polychlorinated biphenyls (PCBs) are ideally performed
with PCB mixtures reflecting the composition of environmental PCB profiles to mimic actual effects
and to account for complex interactions among individual PCB congeners. Unfortunately, only a few
laboratory studies employing synthetic PCB mixtures have been reported, in part because of the
challenges associated with the preparation of complex PCB mixtures containing many individual
PCB congeners. The objective of this study was to develop a PCB mixture that resembles the average
PCB profile recorded from 1996 to 2002 at a satellite station of the Integrated Atmospheric
Deposition Network located at the Illinois Institute of Technology (IIT) in Chicago, Illinois, using
commercial PCB mixtures. Initial simulations, using published Aroclor profiles, showed that a
mixture containing 65% Aroclor 1242 and 35% Aroclor 1254 was a good approximation of the target
profile. A synthetic Chicago air mixture (CAM) was prepared by mixing the respective Aroclor's in
this ratio, followed by GC/MS/MS analysis. Comparison of the PCB profile of the synthetic mixture
with the target profile suggests that the synthetic PCB mixture is a good approximation of the average
IIT Chicago air profiles (similarity coefficient cos = 0.82; average relative percent difference =
84%). The synthetic CAM was also a reasonable approximation of the average of 184 PCB profiles
analyzed in 2007 at 37 sites throughout Chicago as part of the University of Iowa Superfund Basic
Research Program (isbrp), with a cos of 0.70 and an average relative percent difference of 118%.
While the CAM and the two Chicago air profiles contained primarily di- to pentachlorobiphenyls,
higher chlorinated congeners, including congeners with seven or eight chlorine atoms, were
underrepresented in the synthetic CAM. The calculated TCDD toxic equivalency quotients of the
synthetic CAM (2.7 ng/mg PCB) and the IIT Chicago air profile (1.6 ng/mg PCB) were comparable,
but lower by two orders of magnitude than the isbrp Chicago air profile (865 ng/mg PCB) due to
surprisingly high PCB 126 levels in Chicago air. In contrast, the calculated neurotoxic equivalency
quotients of the CAM (0.33 mg/mg PCB) and the two Chicago air profiles (0.44 and 0.30 mg/mg
PCB, respectively) were similar. This study demonstrates the challenges and methods of creating
and characterizing synthetic, environmental mixtures of PCBs.

*
Corresponding author: Hans-Joachim Lehmler, 100 Oakdale Campus, #221 IREH, Iowa City, IA, Phone: +1-319-335-4211, Fax:
+1-319-335-4290, hans-joachim-lehmler@uiowa.edu.
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Zhao et al.
Page 2
Keywords
Airborne PCBs; Aroclor; PCB homologue; Toxic Equivalency Quotient; TEQ; Neurotoxic
Equivalency Quotient; NEQ; PCB atropisomers
1. Introduction
Congener profiles of polychlorinated biphenyls (PCBs) found in environmental samples are

complex and frequently do not reflect the composition of commercial PCB formulations.
Depending on their three-dimensional structure and their substitution pattern, PCB congeners
bind to different cellular target sites and cause adverse effects by a variety of different
mechanisms. For example, PCB congeners with multiple ortho chlorine substituents may bind
to or activate the constitutive androstane receptor (CAR) (Denomme et al., 1983) and/or the
pregnane X receptor (PXR) (Schuetz et al., 1998). Other ortho substituted PCB congeners
interact with both the aryl hydrocarbon (Ah-)receptor and CAR (Parkinson et al., 1983). PCB
congeners with zero ortho-chlorine atoms avidly bind to the Ah-receptor (Bandiera et al.,
1982) and elicit a toxic response mimicking the action of 2,3,7,8-tetrachlorodibenzo-p-dioxin
(TCDD) (Goldstein et al., 1978; Safe, 1994). In addition, PCB congeners with multiple
ortho substituents sensitize Ryanodine receptors (RyR) (Pessah et al., 2006), an interaction
that has been shown to be enantiospecific in the case of PCB 136 (Pessah et al., 2009). Aside
from receptor-mediated responses, lower chlorinated PCB congeners may be metabolically
activated in vitro and in vivo to reactive intermediates, such as PCB epoxides, quinones and
hydroquinones (Robertson and Gupta, 2000), which in turn can react with cellular targets or
result in the formation of reactive oxygen species (Arif et al., 2003; Lin et al., 2000; Robertson
and Gupta, 2000; Zhao et al., 2004). Primary metabolites of lower chlorinated PCBs may have
a high affinity for the estrogen receptor (ER) (Korach et al., 1988) and, thus, may cause
estrogenic effects.
The complex composition of environmental PCB mixtures and the different modes of action
represent a significant challenge for both environmental and toxicity studies, partly because
the congeners present in PCB mixtures can interact in complex ways to produce effects that
are not apparent from single congener studies. In order to assess health risks in human
populations, it is of particular importance to employ PCB mixtures resembling actual human
exposures for toxicity studies. However, only few toxicity studies with environmentally
relevant PCB mixtures have been reported and most have focused on the ingestion route of
exposure and higher molecular weight congeners. For example, animals have been exposed to
PCB-contaminated soils (Fouchecourt et al., 1998; Hansen et al., 1981b), extracts of soils
(Kania-Korwel et al., 2005; Li and Hansen, 1996b) or pooled extracts of multiple high volume
air samples (Li and Hansen, 1996a). Feeding Great Lakes fish to test animals was used to
closely mimic this important exposure source (Restum et al., 1998; Shipp et al., 1998). Two
studies reported by Larry Hansen and colleagues used fat from Aroclor-exposed animals to
treat another species with the resulting PCB mixture (Hansen et al., 1981a; Hansen et al.,
1983). Several other studies used analyses of exposure sources to formulate synthetic mixtures
from individual PCB congeners. The advantage of this approach is a well-defined PCB mixture,
but environmental mixtures prepared from individual congeners are inevitably incomplete
(Altmann et al., 2001; Gyorkos et al., 1985; Hany et al., 1999; Lilienthal et al., 2000; Parkinson
et al., 1980) and contain only major PCB congeners. A recent study reported the preparation
of a PCB mixture resembling the PCB profile found in Fox river fish using commercial PCB
mixtures (Kostyniak et al., 2005). This approach allows the preparation of large quantities of
a well characterized synthetic mixture containing minor as well as major PCB congeners.
Zhao et al.
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The present study describes the development of a PCB mixture that resembles the average PCB
profile found in Chicago air using two commercial Aroclor mixtures. This approach was
selected because it is very cumbersome to collect adequate quantities of airborne PCB mixtures,
even for short term pilot studies (Li and Hansen, 1996a), and it is simply not feasible to collect
sufficient PCB quantities from air to obtain an environmental mixture for definite studies. The
synthetic mixture was analyzed by GC/MS/MS and compared to two average PCB air profiles
from Chicago. Finally, the PCB homologue composition, the contributions of congeners to the
TCDD and neuro-toxic equivalency quotients TEQ and NEQ and the distribution of chiral PCB
congeners were determined from the congener profile to guide the toxicity assessment of this
synthetic mixture.
2. Materials and methods

2.1. Determination of the average PCB profile observed in Chicago air
A representative, average PCB congener profile in Chicago air was first determined using an
extensive set of airborne PCB congener profiles recorded at the satellite station of the Integrated
Atmospheric Deposition Network (IADN) located at the Illinois Institute of Technology (IIT)
in Chicago, Illinois (Sun et al., 2006). To determine an average congener profile, all the
congener concentrations were normalized as a fraction of the total PCBs. That is, for each
sample the congener concentration (pg/m3) was divided by the sum of the congener
concentrations (Equation 1). PCB is defined as the sum of the concentrations of 93 congeners
or coeluting congener groups.
(Equation 1)
This was repeated for each of the 188 samples reported by IADN for the dates January 12,
1996 through Dec 9, 2002. The average congener fractions for these 188 samples were
considered to be the target profile. To allow a comparison with our analytical data (see below),
this IIT Chicago air profile was collapsed to a profile with 88 congeners or coeluting congeners
groups representing 123 PCB congeners.
Recently, a second large data set with airborne PCB congener profiles became available. As
part of the University of Iowa Superfund Basic Research Program (isbrp), a total of 184 PCB
congener profiles were collected using an innovative sampling strategy of vehicle-mounted
high-volume air samplers at more than 37 sites throughout Chicago, Illinois, in 2007 (Hu et
al., 2008). In the following, the average profile obtained from these individual profiles will be
referred to as the isbrp Chicago air profile.
2.2. Simulation of the synthetic Chicago Air Mixture (CAM)
Initial simulations based on detailed Aroclor compositions reported by George Frame and
colleagues (Cochran and Frame, 1999; Frame, 2001; Frame et al., 1996a; Frame et al.,
1996b; Hansen, 1999) and, subsequently, our own GC/MS/MS analyses (Figures 2 and 3) were
performed to determine which combination of Aroclor 1242 and different Aroclor 1254 would
give the best approximation of the average PCB profile observed in Chicago air. The
contributions from each Aroclor were determined by the method of least squares. That is, the
sum of the squares of the residuals (SSR) between the Chicago profile is minimized to
determine the fraction of the mixture made up from Aroclor 1242 (x) and the fraction made up
from Aroclor 1254 (y)
Zhao et al.
Page 4
where i refers to one of the 92 congeners considered. The terms fi,1242, fi,1254 and fi, IIT refer
to the mass fraction of the total PCBs for each congener i in Aroclor 1242, Aroclor 1254 and
the IIT congener profile, respectively. Since this is a binary mixture,
Microsoft Excel's Solver function was used to perform the iterative calculation and determine
the value of x that would give the minimum SSR value. Using this approach, the best match
of the Chicago air PCB profile was found to be a mixture of 65% Aroclor 1242 and 35% of a
late-production Aroclor 1254 (Johnson et al., 2008). A slightly better approximation of the
target profile could be achieved using a more typical, early production Aroclor 1254 lot.
Unfortunately, large quantities of such an Aroclor 1254 lot were not available for this study.
The average molecular weight of this mixture is 284 g/mol, as calculated from published
molecular weights of Aroclor 1242 (261 g/mol) and Aroclor 1254 (327 g/mol) (Silberhorn et
al., 1990).
2.3. Preparation of the synthetic CAM
Aroclor 1242 (Electrical Grade, Monsanto Lot KB-05-415) and Aroclor 1254 (Electrical
Grade, Monsanto Lot KB-05-612) were generous gifts of Dr. Larry G. Hansen (University of
Illinois, Urbana, Illinois, US). The Aroclor 1254 lot is a well characterized batch and contains
a relatively high percentage of dioxin-like PCB congeners (Johnson et al., 2008). Both Aroclor
lots, in addition to Aroclor's 1248 and 1260, have previously been employed to prepare a PCB
mixture approximating a Fox River PCB profile (Kostyniak et al., 2005). Appropriate amounts
of each Aroclor mixture were weighed into a cleaned and tared brown glass bottle to give 20
grams of a 65:35 mixture. The mixture was shaken continuously for 30 days and small aliquots
were removed after 8, 14 and 30 days for congener specific PCB analysis. The PCB profiles
for all three time points were in excellent agreement with each other.
2.4. GC/MS/MS analysis of the CAM and technical Aroclor mixtures
The quantification of PCB homologues was performed by GC/MS/MS with an Agilent 6890N
gas chromatograph coupled to a Waters Micromass Quattro micro GC mass spectrometer
(Milford, MA, USA) operating under electron impact (EI) positive mode at 70 eV and multiple
reaction monitoring, and the trap current was 200 A as described previously (Hu et al.,
2008). The GC was equipped with an Agilent 7683 series autosampler. Two microlitres of the
sample were injected in splitless mode in a Supelco SPB-Octyl fused silica capillary column
(30 m 250 m i.d. 0.25 m film thickness) with helium as the carrier gas at a constant flow
rate of 0.8 ml min1. The collision gas was ultra pure carrier grade Argon. The initial oven
temperature was 75 C which was held for 5 min. The gradient program ran to 150 C at 15
C/min and was held for 1 min. The second step of the gradient ran to 280 C at 2.5 C/min and
was held for 3 min. A representative GC/MS/MS chromatogram of the mixture obtained under
these conditions is shown in Figure 1. A chromatogram of the PCB standard is included in the
Supporting Material (Figure S1). A total of 182 peaks corresponding to all 209 congeners were
analyzed and the relative amount of each congener was normalized as a fraction of the total
PCB mass in the sample as described above. The congener profile of Aroclor 1254 (i.e., lot
KB-05-612) obtained with this method is in good agreement with the published PCB profile
Zhao et al.
Page 5
of this lot obtained using the same GC column (similarity coefficient cos = 0.92; average
relative percent difference = 75) (Johnson et al., 2008).
3. Results and discussion

3.1. Comparison of the synthetic PCB congener profile with the IIT Chicago air profile
The concentrations and PCB congener profiles in Chicago air are well understood for the
satellite stations of the IADN located at IIT in Chicago. This station has been operating since
1996, collecting samples every 12 days. The average PCB congener distribution signal for this
IADN satellite station was selected as a target profile and is shown in Figure 2A. The objective
of this study was to prepare a synthetic PCB mixture that resembles this Chicago air profile
for further environmental and toxicity studies. Towards this goal, a PCB mixture was simulated
using both detailed PCB congener profiles of Aroclor 1242 and 1254. A mixture containing
65% Aroclor 1242 and 35% Aroclor 1254 gave the best approximation, as determined by the
method of least squares. This synthetic Chicago air mixture (CAM) was prepared by mixing
the respective Aroclor's in a 65:35 ratio followed by thorough mixing for up to 30 days and
analyzed by GC/MS/MS. A congener profile of this synthetic mixture with 88 PCB congeners
or coeluting congener groups representing 123 congeners is shown in Figure 2B.
The differences in the percentage of each congener between the IIT Chicago air profile (Figure
2A) and the synthetic CAM (Figure 2B) are shown in Figure 2C. Although the differences in
percentages were comparatively small, some congeners (e.g., PCB 45 and 118) were clearly
overrepresented in the CAM, whereas other congeners were underrepresented (e.g., PCBs 33,
47&44, 100&95 and 110). This suggests that the IIT Chicago air profile is not a simple mixture
of Aroclor 1242 and Aroclor 1254. This is not a surprising observation because PCBs undergo
congener selective weathering in the environment (Hornbuckle et al., 2006).
One drawback of the IIT Chicago air profile (Figure 2A) is the fact that this profile consists
only of a comparatively small number of PCB congeners. In particular, levels of
monochlorinated PCB congeners are not included in the IIT Chicago air profile. However,
these lower chlorinated PCB congeners are of particular interest because they can be
metabolically activated to reactive and potentially toxic PCB metabolites, such as PCB
epoxides, hydroquinones, semiquinone free radicals and quinones (Amaro et al., 1996; McLean
et al., 1996; Song et al., 2008).
The overall similarity of the IIT Chicago air profile (Figure 2A) and the synthetic CAM (Figure
2B) was further assessed using the similarity coefficient cos (Magar et al., 2005) and the
average relative percent difference (RPD) (Kostyniak et al., 2005). The similarity coefficient
cos , determined using the PCB profiles shown in Figures 2A and 2B, was 0.82 (cos = 1
represents a perfect match). The average RPD, calculated for the RPD of all 88 congeners and
coeluting congener groups was 84%. The RPD has been used previously to determine the
similarity of a synthetic PCB mixture and PCB profile of fish from the Fox River (Kostyniak
et al., 2005). In this study, the average RPD between the two congener profiles was 71% (64
peaks representing 92 PCB congeners). Considering that our study analyzed a larger number
of peaks (88 peaks representing 123 congeners), the RPD of 84% in our study still suggests
that the synthetic CAM is a good approximation of the IIT Chicago air profile.
3.2. Comparison of the synthetic PCB congener profile with the isbrp Chicago air profile
A second dataset with 184 PCB congener profiles collected in 2007 at over 37 locations in
Chicago, Illinois, became recently available (Hu et al., 2008). The PCB profiles of this dataset
were obtained using the same GC/MS/MS method and laboratory used to analyze the synthetic
CAM, which allows a better comparison of the PCB profiles. In addition, this new dataset
Zhao et al.
Page 6
includes highly volatile, monochlorinated PCB congeners, which are frequently not analyzed
in air samples due to analytical challenges. However, these monochlorinated PCB congeners
are readily metabolized to reactive metabolites (Amaro et al., 1996; McLean et al., 1996) that
have been implicated in carcinogenesis (Ludewig et al., 2008).
The average PCB profile showing 169 peaks representing all 209 PCB congeners is shown in
Figure 3A. The analogous PCB profile of the CAM and the difference in the percentage
between both profiles are shown in Figures 3B and 3C, respectively. Several PCB congeners
were over- (e.g., PCBs 5, 45, 66 and 118) or underrepresented (e.g., PCBs 18&30, 20&28 and
68) in the CAM in comparison to the isbrp Chicago air profile in Figure 3A. PCB congeners
underrepresented also include PCB 11, which is typically not found in technical Aroclor
mixtures (Hu et al., 2008), and PCB 126, a highly toxic PCB congener (see below).
The CAM was also a reasonable approximation of the isbrp profile, with a similarity coefficient
cos of 0.70 and an average RPD of 118%. As shown in Figure 3, the CAM was missing in
particular higher chlorinated PCB congeners, which contributes to the comparatively small cos
and the large average RPD. The smaller extent of similarity is in part due to much larger
number of PCB congeners used in the comparison of the PCB profiles.
3.3. Comparison of the homologue composition
Figures 3A-3C show a comparison of the homologue composition of the CAM compared to
the two Aroclor mixtures used in its preparation. The homologue composition of the average
Chicago air profile at the IIT site (Figure 4D) and the average PCB profile recorded at different
locations in Chicago as part of the isbrp (Figure 4E) are shown for comparison. While Aroclor
1254 was a medium chlorinated PCB mixture containing over 50% pentachlorobiphenyls,
Aroclor 1242 contained only a small percentage of pentachlorobiphenyls (6 %). Instead,
Aroclor 1242 was composed of comparable amounts of di-, tri- and tetrachlorobiphenyls
(22-36%). This homologue composition is in agreement with the average molecular
composition reported for these Aroclors (Silberhorn et al., 1990). Because the CAM is a 65:35
mixture of two Aroclor mixtures, it contained a much broader range of homologue groups,
with di- to pentachlorobiphenyls making up significant percentages of this synthetic mixture
(15-34%).
Similar to the CAM, the IIT and isbrp Chicago air profiles were composed of high percentages
of di- to pentachlorobiphenyls (Figures 4D and 4E). However, the IIT Chicago air profile
contained twice as many hepta chlorinated PCB congeners compared to the synthetic CAM
(1.8 versus 0.8 %), i.e. these more persistent, higher chlorinated congeners were
underrepresented in the synthetic mixture. Higher chlorinated PCB congeners with seven or
eight chlorine atoms represented an even more significant percentage of the isbrp Chicago air
profiles (13%). This observation is surprising considering the low vapor pressure of higher
chlorinated PCB congeners (Hornbuckle et al., 2006).
3.4. Comparison of the TCDD Toxic Equivalence Quotient (TEQ)
The number of ortho chlorine substituents determines the preferred conformation adopted by
the two phenyl rings of a PCB congener (Lehmler et al., 2002; Shaikh et al., 2008) and, thus,
determines its cellular targets. Several congeners with zero or one ortho chlorine atom can
adopt a conformation similar to TCDD and interact with the aryl hydrocarbon (Ah)-receptor
(Bandiera et al., 1982). These PCB congeners display dioxin-like toxicity and have been
assigned a TCDD toxic equivalent factor (TEF) (Van den Berg et al., 2006). Nine dioxin-like
PCB congeners, including PCBs 77, 81, 105, 114, 118, 123, 156, 157 and 167, were detected
in the three synthetic mixtures, whereas PCBs 126, 167 and 169 were below the detection limit
(Figure 5A-5C). The observation that PCB 126 was not detected is surprising because the
Zhao et al.
Page 7
Aroclor 1254 lot used in this study has been reported to resemble the 1254 A lot (Kostyniak
et al., 2005) described by Frame et al. (Frame, 2001). Specifically, the Aroclor 1254 A lot
contains 0.02% of PCB 126 and, thus, has a high TEQ activity compared to other Aroclor 1254
lots.
A theoretical TEQ based on the 2005 WHO TEF (Van den Berg et al., 2006) was calculated
for the two Aroclors and the CAM by multiplying the weight percentage with the TEF of each
congener. The TEQs the sum of these products are summarized in Table 1 and decreased
in the order TEQAroclor 1254 > TEQCAM > TEQAroclor 1242. PCB 118, which was a major
constituent of the three synthetic PCB mixtures, made the most significant overall contribution
to the TEQ, followed by PCBs 105 and 77. The TEQ of the Chicago air profile determined at
the IIT site appeared to be comparable to the synthetic PCB mixtures (Figure 5D). However,
in contrast to the three synthetic PCB mixtures, PCBs 81 followed by PCBs 118 and 123 made
major contributions to the TEQ of this Chicago air profile (PCB 105 was not included in the
calculation of the TEQ because it co-eluted with PCBs 153 and 132).
The TEQ of the isbrp Chicago air profile was 865 ng/mg PCB as compared to 2.69 ng/mg PCB
for the synthetic CAM and 1.59 ng/mg PCB for the IIT Chicago air profile (Table 1). Although
all eleven PCB congeners with a TEF were reported for this profile, PCB 126 and, to a lesser
extent, PCB 169 made the almost exclusive contributions to the TEQ of this profile (Figure
5E). Both PCB 126 and 169 have a high affinity for the Ah-receptor and, therefore, large TEF
values (Van den Berg et al., 2006). However, the apparently high TEQ of the isbrp Chicago
air profile needs to be interpreted with caution because PCBs 126 and 169 were detected only
at 7% and 10% of the sampling sites.
3.5. Comparison of the Neurotoxic Equivalence Quotient (NEQ)
In contrast to dioxin-like PCB congeners, congeners with two or more ortho chlorine
substituents adopt conformations with larger dihedral angles than dioxin like PCB congeners
and, therefore, do not interact with the Ah-receptor (Bandiera et al., 1982). As discussed above,
these non-dioxin like PCB congeners cause adverse effects, such as (developmental)
neurotoxicity, by other modes of action. Simon et al. recently proposed a neurotoxic
equivalence scheme of relative potency of PCB congeners (Simon et al., 2007). The NEQs
calculated for all five PCB profiles using the neurotoxic equivalent values (NEV) estimated
by Simon et al. are very similar (Table 1). In contrast to the TEQ, a large number of PCB
congeners (52-64 out of 74 congeners with a NEV), mostly congeners with two to five chlorine
substituents, made minor contributions (i.e., < 5%) to the NEQ (Figure 6). PCB 45 was a major
neurotoxic PCB congener in the three synthetic PCB mixtures (7.7 to 14.3 % NEQ; Figure
6A-6C), but made only a minor contribution to the NEQ of the IIT and the isbrp Chicago air
profiles (< 2 % NEQ; Figure 6D and 6E). PCB 52 and 95 made significant contributions to the
NEQ of the IIT and the isbrp Chicago air profiles (> 15 % NEQ). While the contribution of
PCB 52 to the NEQ in the CAM was comparable to its role in the two Chicago air profiles,
PCB 95 was underrepresented in the CAM based on its contribution to the NEQ.
3.6. Comparison of the relative composition of chiral PCB congeners
Some neurotoxic PCB congeners with three or four ortho chlorine substituents and
unsymmetrical substitution patterns in both phenyl rings are chiral due to the hindered rotation
around the biphenyl bond and, like other chiral organic compounds, can be subject to
enantioselective disposition processes or interact with cellular targets in an enantio-selective
or specific manner (Lehmler and Robertson, 2001; Pessah et al., 2009). Figures 7A-C show
that the congener distribution of chiral PCBs differs among the three synthetic mixtures. All
three synthetic mixtures contain comparable total amounts of chiral PCBs ranging from 7.2 to
8.3 %, with PCBs 45, 84, 95 and 132 comprising about 6.3 to 6.7 % of the respective mixture.
Zhao et al.
Page 8
However, the overall distribution of chiral PCB congeners changes significantly with
increasing degree of chlorination of the three synthetic mixtures. For example, the percentage
of PCB 45 decreases from 5.9 % (80% of chiral PCBs) in Aroclor 1242 to 2.2 % (27 % of
chiral PCBs) in Aroclor 1254. Furthermore, Aroclor 1254 contains traces of many higher
chlorinated, chiral PCBs, such as PCBs 88, 91, 135, 136, 144, 149, 171, 174 and 183. Chiral
PCBs 131, 175, 176, 196 and 197 were not detected in any of the PCB mixtures. In contrast,
the two Chicago air profiles display a different composition with regards to chiral PCB
congeners (Figures 7D and 7E). Specifically, the neurotoxic PCB 95 is a major chiral PCB
congener in both Chicago air profiles, whereas PCB 45 represents only a comparatively small
percentage of chiral PCB congeners.
Supplementary Material
Refer to Web version on PubMed Central for supplementary material.
Acknowledgments
This research was supported by grants ES05605, ES012475 and ES013661 from the National Institute of
Environmental Health Sciences (NIEHS), NIH, and Major Research Instrumentation grant BES-0420378 from the
National Science Foundation. Contents of this manuscript are solely the reponsibility of the authors and do not
necessarily represent the official views of NIEHS and NSF.
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Figure 1.
Representative GC/MS/MS chromatograms for the synthetic Chicago air mixture (CAM)
consisting of 65% Aroclor 1242 and 35% Aroclor 1254 showing the homologue composition.
The analysis was done in the monitored reaction mode. A detailed description of the gas
chromatographic separation is provided under Materials and Methods.

Zhao et al.
Page 13

Figure 2.
Comparison to the synthetic CAM with the average PCB profile in Chicago air. (A) Average
PCB IIT Chicago air profile; (B) PCB profile of the synthetic CAM consisting of 65% Aroclor
1242 and 35% Aroclor 1254; (C) difference in percentage of the PCB congener profiles
(synthetic CAM minus the IIT Chicago air profile).
Zhao et al.
Page 14

Figure 3.
Comparison of the average 2007 PCB profile in Chicago air with the synthetic CAM. (A)
Average PCB isbrp Chicago air profile; (B) PCB profile of the synthetic CAM consisting of
65% Aroclor 1242 and 35% Aroclor 1254; (C) difference in percentage of the PCB congener
profiles of both mixtures (synthetic CAM minus the isbrp Chicago air profile). The labels on
the x-axis show only every other PCB congeners.
Zhao et al.
Page 15

Figure 4.
Homologue composition of (A) Aroclor 1242, (B) Aroclor 1254, (C) the synthetic CAM, (D)
the average IIT Chicago air profile and (E) the average isbrp Chicago air profile.

Zhao et al.
Page 16

Figure 5.
Relative contribution of dioxin-like PCB congeners to the TCCD Toxic Equivalency Quotient
(TEQ) of A) Aroclor 1242, (B) Aroclor 1254, (C) the synthetic CAM, (D) the average IIT
Chicago air profile and (E) the average isbrp Chicago air profile. TEQ values were calculated
from the PCB congener profiles using the 2005 WHO TCDD toxic equivalent factors (TEF)
(Van den Berg et al., 2006). PCB 105 was not included in the TEQ calculation for the IIT
Chicago air profile because it co-eluted with PCBs 153 and 132.

Zhao et al.
Page 17

Figure 6.
Relative contribution of PCB congeners with a Neurotoxic Equivalency Factor (NEF) to the
Neurotoxic Equivalency Quotient (NEQ) of A) Aroclor 1242, (B) Aroclor 1254, (C) the
synthetic CAM, (D) the average IIT Chicago air profile and (E) the average isbrp Chicago air
profile. The NEQ for each mixture was calculated from the PCB congener profiles using the
neurotoxic equivalence factors proposed by Simon et al. (Simon et al., 2007).

Zhao et al.
Page 18

Figure 7.
Weight percentage of chiral PCB congeners in (A) Aroclor 1242, (B) Aroclor 1254, (C) the
synthetic CAM, (D) the average IIT Chicago air profile and (E) the average isbrp Chicago air
profile. Only major chiral PCB congeners are shown.

Zhao et al.
Page 19
Table 1
Comparison of the TCCD Toxic Equivalency Quotient (TEQ) and Neurotoxic Equivalency Quotient (NEQ) of
the PCB profiles.
PCB mixture
TEQ [ng/mg PCB]
NEQ [mg/mg PCB]
Aroclor 1242
0.5
0.36
Aroclor 1254
7.9
0.26
CAM
2.7
0.33
IIT Chicago air profile
1.6
0.44
865a
0.30
isbrp Chicago air profile

a
PCBs 126 and 169 were detected only at 7% and 10% of the sampling sites.

Environ. Sci. Technol. 2010, 44, 28222827
Inadvertent Polychlorinated
Biphenyls in Commercial Paint
Pigments
DINGFEI HU AND KERI C. HORNBUCKLE*
Department of Civil & Environmental Engineering and
IIHR-Hydroscience and Engineering, The University of Iowa,
Iowa City, Iowa 52242
Received August 7, 2009. Revised manuscript received

September 17, 2009. Accepted November 16, 2009.
A polychlorinated biphenyl (PCB) that was not produced as

part of the Aroclor mixtures banned in the 1980s was recently
reported in air samples collected in Chicago, Philadelphia,
the Arctic, and several sites around the Great Lakes. In Chicago,
the congener 3,3-dichlorobiphenyl or PCB11 was found to
be the fifth most concentrated congener and ubiquitous throughout
the city. The congener exhibited strong seasonal concentration
trends that suggest volatilization of this compound from
common outdoor surfaces. Due to these findings and also the
compounds presence in waters that received waste from
paint manufacturing facilities, we hypothesized that PCB11 may
be present in current commercial paint. In this study we
measured PCBs in paint sold on the current retail market. We
tested 33 commercial paint pigments purchased from three
local paint stores. The pigment samples were analyzed for all
209 PCB congeners using gas chromatography with tandem
mass spectrometry (GC-MS/MS). More than 50 PCB congeners
including several dioxin-like PCBs were detected, and the
PCB profiles varied due to different types of pigments and
different manufacturing processes. PCB congeners were detected
in azo and phthalocyanine pigments which are commonly
used in paint but also in inks, textiles, paper, cosmetics, leather,
plastics, food and other materials. Our findings suggest
several possible mechanisms for the inadvertent production of
specific PCB congeners during the manufacturing of paint
pigments.
Introduction
Polychlorinated biphenyls (PCBs) are a family of 209 compounds, called congeners, produced commercially as Aroclors
by chlorination of biphenyl. The Aroclor mixtures were
marketed for use in electrical transformers, capacitors, heat
transfer systems, and hydraulic systems (1, 2). Lower quantities were used in voltage regulators, adhesives, caulking
compounds, inks, lubricants, paints, sealants, carbonless
copy paper, coatings, electrical switches, plasticizers, circuit
breakers, dust control agents, and older fluorescent lighting
fixtures (2). Aroclors were used in paint formulations as drying
oils (resins) and plasticizer or softening agents (liquids). Data
provided to EPA indicate that PCBs have been found in dried
paint at concentrations that range from less than 1 ppm to
97,000 ppm (3).
Part of the special section Sources, Exposures, and Toxicities

of PCBs in Humans and the Environment.
* Corresponding author phone: (319) 384-0789; fax: (319) 3355660; e-mail: keri-hornbuckle@uiowa.edu.
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ENVIRONMENTAL SCIENCE & TECHNOLOGY / VOL. 44, NO. 8, 2009
Some PCB congeners, usually called non-Aroclor PCBs,

are not present or are very low in concentration due to
unfavored or improbable formation during the Aroclor
manufacturing process (2). PCB11 is one of such non-Aroclor
PCB congeners. In air samples from Chicago collected in
2007, we found PCB11 widely distributed throughout the
city (4). The compound was almost simultaneously reported
in air of polar regions (5). Since then, its presence was also
reported in air of Philadelphia (6) and five sites around the
Great Lakes (7). It appears that PCB11 is a global pollutant.
In addition, PCB11 was measured in the wastewater effluent
from paint production. Possible production of PCB11 from
dechlorination is not likely because its possible precursors
are in very low concentration in Aroclors (8, 9). The
widespread distribution of PCB11 throughout Chicago and
elsewhere suggests volatilization of this compound from
surfaces. Litten et al. reported that PCB11 was in surface
waters and effluent waste streams from a pigment manufacturing plant around New York Harbor (10). Recently,
Rodenburg et al. detected it in consumer goods including
newspapers, magazines, and cardboard boxes, which usually
contain color pigments (11). Therefore, we hypothesize that
PCB11 and other PCB congeners are present as byproduct
in current commercial pigments.
Paint is composed of pigments, solvents, resins, and
various additives (12). Two major groups of paints are latex
(water-based) and alkyd (oil-based) paints (13). The major
difference between latex and alkyd paints is that the major
liquid portion of latex paints is water while the liquid in
oil-based paints consists of petroleum distillates and other
organic solvents such as toluene and xylene. Latex paints are
the most common type for house use from exterior paint
and trim, to interior walls and woodwork. Generally, a paint
store has about 10 different colors of base pigments, and
paints are sold by mixing pigments with other components.
To test our hypothesis, we purchased and analyzed paint
pigments from three paint stores. According to IBISWorld
Inc., in 2007 these companies account for about 70% of the
market share in the United States.
Materials and Methods

Reagents. Paint pigments were purchased from three different paint retailers: Sherwin Williams, PPG Pittsburgh, and
Vogel, in Iowa City, Iowa in 2009. A calibration standard
solution with a full suite of 209 PCB congeners was prepared
from five PCB congener solutions purchased from AccuStandard (New Haven, CT). Acetone and hexane (pesticide
grade) were purchased from Fisher Scientific (Fair Lawn,
NJ).
Sample Analysis. The extraction method was modified
from U.S. EPA method 3545 (14). In brief, approximately
5.0 g of the fresh pigment sample was accurately weighed
and mixed with combusted diatomaceous earth, then spiked
with 50 L of 500 ng/mL surrogate standards containing
PCB14 (3,5-dichlorobiphenyl), PCB65 (2,3,5,6-tetrachlorobiphenyl) and PCB166 (2,3,4,4,5,6-hexachlorobiphenyl) (Cambridge Isotope Laboratories, Inc.). The samples were extracted
utilizing a pressurized fluid extraction (Accelerated Solvent
Extractor, Dionex ASE-300) with a mixture of acetone and
hexane (1:1, v/v). The extract was concentrated to 2 mL
from 200 mL, and the concentrated extract was transferred
to a glass test tube; 2 mL of concentrated sulfuric acid was
slowly added and mixed. Hexane (8 mL) was used to extract
the acidified mixture 3 times after a 10-min mechanical
shaking and centrifugation at 3000 rpm/min for 5 min. The
pooled extract was concentrated down to 2 mL and passed
10.1021/es902413k
2010 American Chemical Society
Published on Web 12/03/2009
through a silica gel column prepared with 0.1 g of combusted

silica gel at the bottom and 1 g of acidified silica gel (2:1silica
gel:concentrated sulfuric acid by weight). Hexane (10 mL)
was used to elute PCBs from the column and the eluate was
concentrated down to 0.5 mL for PCB analysis. All samples
were analyzed in duplicate, and the average is reported.
The final extract was spiked with 20 ng of PCB204
(2,2,3,4,4,5,6,6-octachlorobiphenyl) as internal standard
(Cambridge Isotope Laboratories, Inc.). All 209 PCB congeners, in about 170 chromatographic peaks, were analyzed
using a gas chromatograph with mass selective detection
(GC-MS/MS) modified from the EPA method 1668A (15). The
quantification of PCB congeners was performed by an Agilent
6890N gas chromatograph with an Agilent 7683 series
autosampler coupled to a Waters Micromass Quattro micro
GC mass spectrometer (Milford, MA) operating under
electron impact (EI) positive mode at 70 eV and multiple
reaction monitoring (MRM), and the trap current was 200
A. The retention windows were defined by PCB parent/
daughter ion pairs from mono- to deca- homologues which
were 188/152, 222/152.10, 255.96/186, 291.92/222, 325.88/
255.90, 359.84/289.90, 393.80/323.90, 427.76/357.80, 461.72/
391.83, 497.68/427.70, respectively.
Quality Assurance/Quality Control. During extraction
of paint pigments using ASE, samples and blanks (combusted
diatomaceous earth) were alternated on the instrument to
avoid, and detect, any cross contamination between pigment
samples. The average recoveries of PCB 14, PCB 65, and PCB
166 surrogate standards injected in every sample were 56 (
15%, 85 ( 25%, and 86 ( 20%, respectively. Di- to tri- PCB
concentrations in samples were corrected for PCB14 recovery
and tetra- to deca-PCBs for PCB166 based on our analytical
method validation using Standard Reference Material 1944.
Results and Discussion

Inorganic and Organic Pigments. Inorganic pigments are
produced from either naturally mined pigments (sienna,
umber, ochre) or synthetically manufactured pigments (iron
oxide, carbon black, etc). Titanium dioxide is the most
important white pigment in the industry because of its high
refractive index, reflectance, ease of dispersion, brightness,
and opacity (16, 17). Titanium dioxides and iron oxides
account for approximately 70% and 15% of world consumption of inorganic pigments (18, 19). No PCBs were found in
inorganic pigments which primarily contain titanium dioxide,
iron oxide, raw umber, or carbon black (Figure 1 and Table
1). PCB formation is expected to be associated with chlorinated solvent or intermediates used in the manufacturing
process of pigments.
PCBs were primarily found in organic paint pigments with
a concentration range from 2 to 200 ng/g fresh weight (f.w.)
in 15 of 33 analyzed paint pigments (Figure 1 and Table 1)
in this study. Pigment chemical structures were provided by
Sherwin Williams. PPG Pittsburgh and Vogel did not provide
this information, although we were able to determine several
pigment types based on their material safety data sheets.
Most orange, red, and yellow pigments are made from azo
pigments, and PCBs are only found in two groups of organic
pigments: azo pigments and phthalocyanine pigments. For
pigment samples from Sherwin Williams, we clearly see PCBs
are only present in these two types of pigment. Chlorinated
solvents or intermediates are usually involved to produce
these two types of organic pigments, and side-reactions of
these chlorinated compounds result in formation of PCBs
during the manufacturing process. The EPA is aware of the
presence of PCBs in diarylide pigments and phthalocyanine
pigments. Diarylide pigments belong to the azo category of
pigments (20). However, we observed the presence of PCBs
not only in diarylide pigments but also in other azo pigments
FIGURE 1. PCB concentrations in 33 commercial paint

pigments purchased from Sherwin Williams, PPG Pittsburgh,
and Vogel paint stores.
such as Hansa yellow, quinacridone, isoindolinone, and
maybe more, since some pigment types are unknown.
Azo and phthalocyanine pigments and chemically identical dyes are the most important groups of synthetic colorants
with a great variety of industrial applications. They are used
for coloring paints, inks, textiles, paper, cosmetics, leather,
plastics, food and other materials (21, 29). The widespread
use of these pigments explains the presence of PCB11 in
commercial goods common throughout modern society, such
as newspapers, magazines, and cardboard boxes (11).
Although we do not know if inadvertent PCBs have adverse
effects on human health, there are many potential routes for
human exposure to these PCBs through inhalation, dermal
exposure, and ingestion due to their physicochemical
characteristics of semivolatility, hydrophobicity, and persistence.
Congener Profiles. The detailed PCB distribution profile
in each pigment is provided in Tables S1-S3, and two
examples are presented in Figure 2. The pigments, Y1 of
Sherwin Williams and 96-26Z of PPG Pittsburgh, are both
yellow and made from monoazo yellow pigments. The
synthesis of monoazo yellow pigments involves the coupling
of a diazotized substituted aniline with a coupling component
containing an active methylene moiety in a linear structure
(18). There are different PCB distribution profiles in different
pigments due to various manufacturing processes for different pigments or even the same pigments. The same type
pigment might have different starting materials, intermediates, or manufacturing conditions. For a particular manufacturing process, only very limited numbers of chlorinated
compounds are involved; however, up to 22 congeners were
detected in one pigment. Among these detected congeners,
PCBs 77, 114, and 123 are dioxin-like congeners which have
distinct toxic properties. A variety of PCB profiles in paint
pigments were observed in this study (Tables S1-S3),
although the reason for their presence is not completely
understood. PCB11 was most often detected: it was found in
13 of 15 pigment samples for which any PCBs were detected,
followed by PCBs 8, 6, 4, 1, 12/13, 2, 3, and 209seach with
more than 40% detection frequency (Figure 2). PCB congeners
of all chlorination levels were found in the pigments.
VOL. 44, NO. 8, 2009 / ENVIRONMENTAL SCIENCE & TECHNOLOGY
2823
TABLE 1. Colors and Types of Commercial Paint Pigments Purchased from Three Paint Storesa
paint store
code
pigment type
Sherwin Williams
Y1
G2
R4
L1
W1
N1
Y3
R2
R3
B1
yellow
green
red
blue
white
raw umber
deep gold
maroon
magenta
black
hansa yellow
phthalocyanine green
isoindolinone
phthalocyanine blue
titanium dioxide
raw umber titanium dioxide
iron oxide
iron oxide
quinacridone
carbon black
PPG
96-5E
96-4D
96-13M
96-10J
96-26Z
96-7G
96-6F
96-12 L
96-23W
96-2B
96-3C
96-22 V
blue
green
durable red
carbazole violet
medium yellow
durable yellow
red
raw umber
white
lamp black
yellow oxide
violet
phthalocyanine blue
/
/
monoazo yellow
/
iron oxide
/
titanium dioxide
/
iron oxide
quinacridone
Vogel
CC
DD
PP
HH
TT
MM
VV
FF
EE
JJ
KK
blue
magenta
green
exterior red
medium yellow
red oxide
white
raw umber
black
yellow oxide
brown oxide
phthalocyanine blue
/
/
/
/
/
/
/
/
/
/: proprietary.
However, lower chlorinated PCB congeners dominated in

most pigment samples except in phthalocyanine green
pigments which contain very high levels of PCB209 relative
to other detected congeners. PCB209 accounts for approximately 66%, 33%, and 50% of total PCBs in phthalocyanine green pigments of G2, 86-4D, and PP, respectively,
from three different paint stores. It is interesting that although
dissimilar in structure, both PCB 11 and PCB 209 are nonAroclors that could be used as signatures of pigment use or
discharge.
Formation Mechanism. Phthalocyanine Pigments. Copper phthalocyanine pigments are the most widely used blue
and green pigments for various applications (16, 21). Two
different general processes are used for commercial production of phthalocyanine pigments: one is from phthalic
anhydride, urea, and copper or a copper salt, and the second
is from phthalonitrile and copper or a copper salt (22). The
first route is less expensive and is usually used to produce
phthalocyanine pigments for high-volume and low-cost
applications such as paint pigments and dyes for textiles
and paper; the second route is more expensive but produces
high-quality and high-purity phthalocyanine pigments such
as charge generation materials for electrophotography (21).
The first urea process usually involves organochlorine solvent
such as di- or trichlorobenzene as the reaction medium. Uyeta
et al. showed that starting materials (urea, phthalic anhydride,
copper chloride, ammonium molybdate) and the initial
reaction medium (di- or trichlorobenzene) did not contain
PCB congeners (22), so they are not a direct source of PCBs
in pigments.
PCB formation mechanisms (Figure 3) are proposed for
the urea manufacturing process. Phthalocyanine blue is
2824
color
produced from starting materials without chlorines, and

phthalocyanine green is derived from phthalocyanine blue
by chlorination (21). Lower chlorinated PCB congeners are
produced as by-products during the manufacturing process
of phthalocyanine blue pigments from the reaction medium
chlorobenzene (23). The reaction medium, dichlorobenzene
or trichlorobenzene, can form tetra-, penta-, and/or hexaPCB congeners by a reaction with each other under heat
through a free radical mechanism (the dashed arrow pathway
in Figure 3) (24, 25). The resulting PCB congeners may
thermally degrade further into lower chlorinated congeners
by the same mechanism (26). Mono- through tetra-chlorobiphenyls have been created through a free radical mechanism from pyrolysis and combustion of other chlorinated
organics at temperatures ranging from 300 to 700 C (27).
The free radical mechanism to form PCBs from chlorobenzenes has been experimentally demonstrated (25). The
temperature is usually below 300 C during the pigment
manufacturing process; however, the presence of copper
chloride and ammonium molybdate as a catalyst might
promote this mechanism at a lower temperature range
(25, 28). To gain proper brightness, shade, strength, and flow
properties of synthetic pigments, various factors including
the reaction temperature and the drying temperature might
be altered to meet these purposes (20). With increase of the
reaction temperature, the total PCB formation increases
independent of reaction time after the initial 2 h (25). Lower
chlorinated benzenes might produce more PCB congeners
than higher ones.
During the process of perchlorination from phthalocyanine blue to phthalocyanine green, decachlorobiphenyl (PCB
209) is formed along with some other highly chlorinated
FIGURE 2. Examples of PCB profiles in paint pigments (top two plots) and the frequency of congener detection in the 15 pigments
with detected PCBs (bottom plot).
FIGURE 3. PCB formation mechanisms in the manufacture process of phthalocyanine blue and phthalocyanine green. The subscripts
x, a, b, and c refer to the number of chlorine atoms.
congeners such as nonachlorobiphenyls (PCBs 206, 207, and
208) from less chlorinated congeners (Figure 3). This explains
the presence of much more nona- and deca-PCB congeners
in phthalocyanine green than in phthalocyanine blue, which
can be observed by comparison of PCB distribution profiles
in L1 and G2 from Sherwin Williams, 96-5E and 96-4D from
PPG Pittsburgh, and CC and PP from Vogel.
Azo Pigments. Azo pigments are the most important group
of synthetic colorants with the largest fraction (more than
50%) of organic pigments on the market (29). Azo pigments

have a wide range of colors covering almost the entire visible
spectrum although blue and green colors are mostly provided
by phthalocyanine and two other pigments (18). Some vivid
colors of azo pigments are commercially dominant, especially
reds, oranges, and yellows.
Azo pigments are almost exclusively produced through a
reaction sequence of diazotization and coupling to afford
the azo group (sNdNs) which is the chromophore responVOL. 44, NO. 8, 2009 / ENVIRONMENTAL SCIENCE & TECHNOLOGY
2825
FIGURE 4. Possible formation pathways of PCB11 and PCB52 in

pigments.
sible for its vivid colors (18). The reaction involves a primary
aromatic amine as a diazo component and a nucleophilic
aromatic or aliphatic compound with active methylene
groups as a coupling component. An aromatic amine such
as a mono-, di, or trichlorinated aniline is typically involved
in the diazotization reaction as a diazo component, and
frequently they are also used as coupling components.
Another important group of diazo components for azo
pigment formation include 3,3-dichlorobenzidine, and to a
lesser extent 2,25,5-tetrachlorobenzidine, 3,3-dimethoxybenzidine, and 3,3-dimethylbenzidine (18). The last two
compounds do not contain chlorines which are required
elements for formation of PCB congeners.
For synthesis of azo pigments, there are more than 10
common intermediates and starting materials such as
chlorinated aniline and chlorinated benzidines that can
potentially have side-reactions to produce PCBs. PCBs are
probably formed by coupling of bis-diazotized dichlorobenzidines or tetrachlorobenzidines under basic conditions as
a result of the decomposition of the diazo moiety. Polychlorinated anilines can be also used to form PCBs through
the free radical mechanism, and the free radical rearrangement of chlorine positions might play a significant role in
varieties of PCB congeners with limited chlorinated intermediates. Lower PCB congeners may be formed by carrying
out the coupling process at lower pH or in the presence of
unsaturated aliphatic compounds such as acylamides (20).
For example, 3,3-dichlorobenzidine and 2,2,5,5-tetrachlorobenzidine are probably the diazo components for TT and
HH pigments that contain high PCB11 and PCB52 (Table
S3), respectively. Unfortunately, we cannot verify the pathways for pigments considered proprietary by manufacturers;
however, the links of intermediates and PCB by-products
can be illustrated structurally (Figure 4). The azo pigments
based on 3,3-dichlorobenzidine appear preponderant (30),
which might explain partially the consistency of PCB11
detection.
Environmental Emission. A wide variety of organic
pigments are commercially available; however, in terms of
chemical structure, almost all currently produced organic
pigments belong to four different groups: azo pigments and
lakes (salt type), phthalocyanine pigments, polycyclic pigments, and heterocyclic pigments (20). In spite of accelerated
progress in the synthesis of organic pigments, commercially
available pigments at present are chemically identical to those
produced historically since the use of synthetic pigments.
PCB congeners are primarily detected in azo pigments and
phthalocyanine pigments. PCB11 is consistently detected in
almost all azo and phthalocyanine pigments, and it is absent
or in very low relative concentrations in commercial Aroclor
mixtures. Therefore, PCB11 can be regarded as a key indicator
of PCB emission from de novo synthesis as by-products of
2826
industrial synthetic process of paint pigments. PCB11 is the

fifth highest congener and ubiquitous in Chicago air (4).
Although we do not know the contribution of PCB congeners
from paint pigments to the airborne PCBs in the environment,
these congeners, especially low chlorinated congeners, might
contribute a significant portion as PCB11 because of their
high volatility.
Based on 40 CFR 761.80, PCBs are allowed at less than 25
mg/kg with a 50 mg/kg maximum in commerce of diarylide
pigments or phthalocyanine pigments when leaving a
manufacturing site or imported to the United States. PCB
levels in the examined paint pigments are all below the
regulatory standard; however, paints are being extensively
and constantly used especially in urban areas. PCBs might
accumulate due to their resistance to degradation in the
environment. It has been reported that PCB11 and total PCB
levels in air are directly proportional to human population
density (7, 31, 32). To our knowledge, pigments or dyes are
the only significant source of PCB11. The elevation of PCB11
in air must be associated with human activity utilizing
pigments or dyes. The presence of PCB11 indicates paint
should be an important source of airborne PCBs although
the link of PCBs in paint pigments and PCBs in air is still not
clear.
Acknowledgments
We thank our analytical core coleader, Craig Just, and our
laboratory director, Collin Just. We also thank Kristin Isley
and Timothy Schulz for assisting with sample preparation in
the laboratory. Funding for this work was provided by the
National Institute for Environmental Health Sciences (NIEHS/
NIH) Superfund Basic Research Program (ES013661).
Supporting Information Available

Three tables. This information is available free of charge via
the Internet at http://pubs.acs.org.
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colorants: influence of solubility and bioavailability. Toxicol.
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Faktor-faktor yang berperan pada karsinogenesis

*)Bagian
Elna Kartawiguna*)
Histologi Fakultas Kedokteran Universitas Trisakti
ABSTRACT
The majority of cancers are caused by extrinsic factors. These include environmental carcinogens
(chemical carcinogen, radiation and virus) and modifying factors (e.g., hormonal imbalance and dietary
deficiency).Genetic and psychological factors also play a role in determining the probability of an individual
developing cancer. Some cancers have one dominant factor as their causative agent. There are three major
stages in tumor development : initiation, promotion and progression. Cancer epidemiology and experimental
carcinogenesis have established that cancer is a disease caused by multiple factors and multistage in its
development. Because the major causative factor of cancer is extrinsic, theoretically most cancers can be
avoided. A rational approach for prevention is the understanding of cancer etiology and its biologic evolution
into progressively more malignant state. (J Kedokter Trisakti 2001;20(1):16-26)
Key words: Cancer, carcinogenesis, causative factors
ABSTRAK
Sebagian besar kanker disebabkan oleh faktor-faktor ekstrinsik,yaitu semua karsinogen lingkungan
(karsinogen kimia, radiasi dan virus) dan faktor-faktor yang mengubah kondisi kesehatan seseorang (misalnya
ketidak-seimbangan hormonal dan kekurangan zat tertentu dalam makanan). Faktor genetik dan faktor
psikogenik juga mempunyai peranan menentukan kemungkinan seseorang untuk menderita kanker. Beberapa
kanker mempunyai satu faktor yang dominan sebagai penyebabnya. Pertumbuhan kanker dapat dibagi dalam tiga
fase utama yaitu fase inisiasi, promosi dan progresi. Dari penyelidikan epidemiologis kanker dan karsinogenesis
eksperimental disimpulkan bahwa kanker merupakan penyakit yang disebabkan banyak faktor dan terjadinya
melalui banyak fase. Secara teoritis sebagian besar kanker dapat dicegah, karena penyebabnya terutama faktor
ekstrinsik. Pendekatan rasional untuk pencegahan adalah pengertian tentang etiologi kanker dan evolusi
biologiknya yang progresif makin lama makin ganas.
Kata kunci: Kanker, karsinogenesis, faktor penyebab
PENDAHULUAN
Penyakit kanker merupakan penyakit
penyebab kematian nomor 2 di Amerika
Serikat. (1,2) Di Indonesia terdapat kecenderungan peningkatan jumlah penderita kanker
dari tahun ke tahun. Penyelidikan selama 20
tahun terakhir menyimpulkan bahwa 60-90%
penyakit
kanker
berhubungan
dengan
lingkungan, sehingga secara teoritis penyakit
kanker dapat dicegah. Lingkungan di sini
berarti semua yang berinteraksi dengan
manusia yaitu bahan-bahan yang dimakan,
diminum, diisap dan dihirup, juga radiasi,
obat-obatan serta aspek-aspek kelakuan
seksual. (1) Dari penyelidikan epidemiologis
dan laboratoris didapatkan bahwa diet
(misalnya banyak lemak, kurang serat dalam

makanan) mempunyai peranan sebesar 35-50%
untuk timbulnya kanker pada saluran
pencernaan, payu dara, endometrium dan
ovarium. (1) Bahan yang diminum, diisap dan
dihirup (misalnya alkohol, tembakau, debu
asbes) berperanan (22-30%) untuk timbulnya
kanker pada paru, orofarings dan esofagus.
Demikian pula radiasi, faktor genetik dan lain
substansi yang belum diketahui. Faktor
psikogenik berperanan untuk timbulnya kanker
karena mempunyai hubungan dengan imunitas
tubuh.
Untuk mengurangi insidens penyakit ini
diperlukan pengetahuan yang lebih luas
J Kedokter Trisakti, Januari-April 2001-Vol.20, No.1 16
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tentang biologi kanker. Karsinogenesis sampai

saat ini diselidiki melalui pendekatan
epidemiologis, percobaan binatang dan kultur
jaringan. (3)
TEORI KARSINOGENESIS
Pada umumnya, kanker timbul karena
paparan terhadap suatu karsinogen secara
berkali-kali dan aditif pada dosis tertentu,
tetapi pada keadaan tertentu dapat juga timbul
dari dosis tunggal karsinogen. (4) Penyebab
kanker dapat satu karsinogen yang sama
misalnya asap rokok (kanker paru), dapat dua
karsinogen yang berlainan misalnya asap
rokok dan debu asbes (kanker paru) (3), asap
rokok dan radiasi sinar X (kanker paru) (1),
asap rokok dan alkohol (kanker orofarings,
larings dan esofagus) (1,5), gen kanker dan
karsinogen lingkungan. (1) Dari penyelidikan
epidemiologis didapatkan bahwa asap rokok
sebagai karsinogen dan debu asbes sebagai kokarsinogen menimbulkan kanker paru lebih
cepat pada pekerja perokok yang menghirup
debu asbes dibandingkan mereka yang
mengisap asap rokok saja, karena kokarsinogen
membantu
karsinogen
menimbulkan kanker lebih efektif. Dari
penyelidikan epidemiologis juga didapatkan
bahwa bahan yang menghambat mekanisme
pertahanan tubuh membantu timbulnya kanker.
Untuk beberapa macam kanker terdapat satu
faktor yang dominan misalnya sinar ultraviolet
yang menimbulkan kanker kulit dan kelainan
kromosom yang menimbulkan retinoblastoma.
Karsinogenesis yang diinduksi karsinogen
kimia atau fisik maupun biologik memerlukan
waktu yang disebut periode laten yaitu waktu
dari pertama kali terpapar suatu karsinogen
sampai terlihat kanker secara klinis. (1,5)
Periode laten dari kebanyakan kanker
seringkali 20 tahun atau lebih. (1) Efek
karsinogen yang lemah dapat tidak terlihat,
sebab periode latennya melampaui masa hidup
seseorang. (4) Karsinogenesis dapat dibagi
dalam tiga fase utama yaitu fase inisiasi,
promosi dan progresi. (5,6)
Fase inisiasi
Fase ini berlangsung cepat. Karsinogen
kimia misalnya golongan alkylating dapat
langsung menyerang tempat dalam molekul
yang banyak elektronnya, disebut karsinogen
nukleofilik.
Karsinogen
golongan lain
misalnya golongan polycyclic aromatic
hydrocarbon sebelum
menyerang dikonversikan (diaktifkan) dulu secara metabolik
(kimiawi) menjadi bentuk defisit elektron yang
disebut karsinogen elektrofilik reaktif.(5)
Tempat yang diserang adalah asam nukleat
(DNA/ RNA) atau protein dalam sel terutama di
atom nitrogen, oksigen dan sulfur. (4) Air dan
glutation juga diserang, dalam beberapa kasus
reaksi ini di-katalisasi oleh enzim seperti
glutathione-S-transferase. Ikatan karsinogen
dengan DNA menghasilkan lesi di materi
genetik. (6) RNA yang berikatan dengan
karsinogen bermodifikasi menjadi DNA yang
dimutasi. (4,5) Karsinogen kimia yang berikatan
dengan DNA disebut genotoksik dan yang
tidak berikatan dengan DNA disebut
epigenetik. (7) Karsinogen genotoksik dapat
juga mempunyai efek epigenetik. Kokarsinogen dan promotor termasuk dalam
karsinogen epigenetik yang menyebabkan
kerusakan jaringan kronis, perubahan sistem
imun tubuh, perubahan hormon atau berikatan
dengan protein yang represif terhadap gen
tertentu. Jadi karsinogen epigenetik dapat
mengubah kondisi lingkungan sehingga fungsi
sebuah gen berubah, bukan strukturnya. Waktu
yang dibutuhkan dari pertama kali sel diserang
karsinogen sampai terbentuk lesi di materi
genetik adalah beberapa menit. (6) Sel berusaha
mengoreksi lesi ini dengan detoksifikasi
kemudian diekskresi atau dapat terjadi
kematian sel atau terjadi reparasi DNA yang
rusak tersebut oleh enzim sel menjadi sel
normal kembali. (5) Karsinogen kimia dapat
didetoksifikasi/
dinon-aktifkan kemudian
diekskresi atau dapat langsung diekskresi.
Tetapi dari proses pengnon-aktifan ini dapat
terbentuk metabolit yang karsinogenik. (5)
Sebelum terjadi reparasi DNA dapat terjadi
replikasi DNA yaitu satu siklus proliferasi sel
yang menyebabkan lesi DNA tersebut menjadi
permanen disebut fiksasi lesi. (5,6) Waktu yang
dibutuhkan dari pertama kali sel diserang
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karsinogen sampai terjadi fiksasi lesi

(terbentuk sel terinisiasi) adalah beberapa hari
(1-2 hari). (6) Replikasi DNA terjadi karena
terdapatnya sel nekrotik sebagai akibat
karsinogen. Replikasi ini dapat diinduksi oleh
lain bahan kimia toksik, bakteri (misalnya
colitis ulcerativa menjadi kanker kolon,
bronkitis kronis menjadi kanker paru pada
perokok), virus, parasit (schistosomiasis di
Afrika menjadi kanker kandung kemih),
defisiensi diet tertentu, hormon dan prosedur
percobaan seperti hepatektomi parsial. (5) Pada
jaringan yang mengalami peradangan atau
sedang berproliferasi (misalnya luka yang
menyembuh) atau jaringan yang berproliferasi
terus menerus (misalnya sumsum tulang, epitel
saluran pencernaan) tanpa terangsang dari
luarpun dapat terjadi replikasi DNA. Pada
peradangan belum diketahui apakah terjadi
akibat peradangan membantu pertumbuhan sel
atau melemahnya daya tahan tubuh. Sel
terinisiasi dapat mengalami kematian (5) , bila
tidak, maka sel dapat masuk ke fase promosi.
Pada akhir fase inisiasi belum terlihat
perubahan histologis dan biokimiawi hanya
terlihat nekrosis sel dengan meningkatnya
proliferasi sel. (6)
Fase promosi
Sel terinisiasi dapat tetap tenang bila tidak
dihidupkan oleh zat yang disebut promotor.
Promotor sendiri tidak dapat menginduksi
perubahan kearah neoplasma sebelum bekerja
pada sel terinisiasi, hal ini telah dibuktikan
pada percobaan binatang. (1,5) Bila promotor
ditambahkan pada sel terinisiasi dalam kultur
jaringan, sel ini akan berproliferasi. Jadi
promotor adalah zat proliferatif. (4) Promosi
adalah proses yang menyebabkan sel terinisiasi
berkembang menjadi sel preneoplasma oleh
stimulus zat lain (promotor). Pada percobaan
binatang dibuktikan terdapat karsinogen kimia
yang bekerja sendiri sebagai inisiator dan
promotor disebut karsinogen komplit. (1)
Dari penyelidikan pada kultur jaringan
diketahui fase ini berlangsung bertahun-tahun
(10 tahun atau lebih) dan reversibel sebelum
terbentuknya sel tumor yang otonom. (5)
Alkohol adalah promotor untuk kanker
orofarings, larings, esofagus dan hati. (1,6)
Alkohol sebagai promotor pada sirosis hepatis

atau kerusakan hati lain dapat menimbulkan
kanker hati.
Promotor lain yaitu DES (diethylstilbestrol)
adalah estrogen sintetis nonsteroid yang pernah
dipakai untuk terapi osteoporosis, pada tahun
1950
menimbulkan
epidemi
kanker
endometrium. (1,6) DES dosis tinggi pernah
digunakan untuk terapi abortus pada tahun
1940-50 menimbulkan kanker vagina dan
serviks pada anak wanita penderita. Suplemen
estrogen untuk terapi gejala menopause yang
digunakan luas pada tahun 1960 an sampai
pertengahan tahun 1970 menimbulkan epidemi
kanker endometrium. Penyelidikan epidemiologis menunjukkan penurunan insidens
kanker ini ke tingkat semula sesuai dengan
penurunan penggunaannya. Terapi estrogen
masih digunakan pada umumnya dengan
periode lebih pendek sehingga timbulnya
kanker endometrium banyak ber-kurang.
Terapi estrogen juga terbukti meningkatkan
risiko terkena kanker payudara tetapi tidak
sejelas kanker endometrium. Terapi estrogen
meningkatkan penyakit kandung empedu yang
merupakan risiko kanker kandung empedu.
Penyelidikan untuk risiko kanker ovarium
mendapatkan hasil yang berlawanan.
Lemak adalah promotor untuk kanker payu
dara, kolon, endometrium, serviks, ovarium,
prostat dan kandung empedu. (1,6) Pada kanker
payu dara, endometrium dan ovarium karena
lemak menaikkan kadar estrogen. Hasil
penyelidikan epidemiologis dan percobaan
binatang tidak konsisten mengenai diet yang
lebih banyak lemak tidak jenuh gandanya dari
lemak jenuh gandanya dapat menaikkan risiko
terkena kanker.
Obat imunosupresif misalnya azathioprine dan
prednison pada penerima transplantasi organ
adalah promotor untuk macam-macam kanker
terutama kanker sumsum tulang, limfoma,
kanker kulit dan sarkoma Kaposi. (1)
Parasit misalnya Clonorchis sinensis adalah
promotor untuk cholangioma (6) dan
Schistosoma haematobium di Afrika untuk
kanker kandung kemih. (1,6) Steroid anabolik
yang biasa digunakan atlit adalah promotor
untuk hepatoma. (6) Obat kontraseptif estrogen
dosis tinggi tanpa progesteron merupakan
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promotor
untuk
hamartoma
(dapat
menyebabkan perdarahan fatal), kanker
endometrium atau adenoma hati. (6) Setelah
dipakai estrogen dosis rendah dikombinasi
dengan progesteron dosis rendah, risiko kanker
menurun. Penyelidikan epidemiologis menunjukkan obat kontraseptif sekarang tidak
menurunkan atau menaikkan risiko terkena
kanker payu dara dan serviks. Terdapat bukti
obat kontraseptif dapat mencegah terjadinya
kanker ovarium karena obat ini mencegah
ovulasi sebagai efek progesteron (anti
estrogen). Teori kelebihan androgen yang
menimbulkan kanker prostat didukung data
epidemiologis bahwa penderita sirosis hepatis
dan orang yang dikastrasi sedikit yang terkena
kanker prostat. Pada binatang percobaan
testosteron sebagai promotor menyebabkan
kanker prostat. (1)
Esterforbol adalah promotor untuk kanker
kulit, paru dan hati. (4) Kurangnya serat dalam
makanan antara lain menyebabkan kontak
dengan karsinogen lebih lama, memudahkan
seseorang terkena kanker kolon. (6) Dari
penyelidikan didapatkan serat dalam makanan
mungkin menurunkan insidens kanker kolon
dengan cara mencegah interaksi asam empedu
dengan enzim bakteri (flora usus) dalam usus
besar, mencegah pengikatan asam empedu
dengan lain bahan kimia yang karsinogenik
dalam feses, mengurangi waktu feses dalam
usus besar dan menaikkan jumlah feses
sehingga menurunkan konsentrasi karsinogen
dalam usus. (1) Di Inggris ditemukan hubungan
terbalik antara serat pentosa dengan kematian
karena kanker kolon tetapi tidak terdapat
hubungan dengan jenis serat lain atau dengan
keseluruhan serat. (1)
Kurangnya vitamin (A, C, beta-karoten dan E)
dan mikronutrien selenium (Se) dalam
makanan memudahkan seseorang terkena
kanker kulit, hati, orofarings, serviks, kandung
kemih, kolon, lambung, esofagus, larings dan
paru. (6) Kemungkinan vitamin-vitamin ini
memproteksi keganasan terutama dalam
bentuk kombinasi. Dalam saluran pencernaan
vitamin E dan C dapat menghalangi ter(1,5)
bentuknya nitrosamine.
Defisiensi
selenium menaikkan efek karsinogenik
karsinogen kimia pada tikus besar terutama
bila diberi diet tinggi lemak tidak jenuh ganda.

(1)
Di Skandinavia Utara ditemukan hubungan
defisiensi zat besi dengan risiko tinggi terkena
kanker farings dan esofagus. Insidens kanker
lambung 4-5 kali lebih tinggi di negara yang
lebih banyak defisiensi zat besinya daripada di
Amerika Serikat. (1) Pada binatang defisiensi
seng mempunyai hubungan dengan kanker
esofagus dan defisiensi seng dapat berinteraksi
dengan alkohol membantu terbentuknya
kanker esofagus. (1) Suplemen asam folat
mencegah terjadinya kanker serviks pada
wanita yang serviksnya abnormal karena
kontraseptif oral. (1) Konsumsi tinggi kalsium
meningkatkan risiko terkena kanker prostat
terutama bila dikonsumsi melebihi 2000 mg/
hari. (7) Kalsium banyak terdapat dalam susu
skim dan rendah lemak.
Sakarin adalah promotor untuk kanker
kandung kemih pada tikus terutama bila
diberikan selama 2 generasi sedangkan pada
manusia belum terbukti promotor untuk
kanker. (1) Penyelidikan epidemiologis
menunjukkan risiko terkena kanker kandung
kemih meningkat 60% pada pria tetapi
penyelidikan lain gagal memastikan kenaikan
ini. (1) Siklamat adalah promotor untuk kanker
kandung kemih pada binatang percobaan
sehingga pada tahun 1969 dilarang peredarannya. (1)
Kopi dihubungkan dengan kanker kandung
kemih dan pankreas pada manusia. Percobaan
kultur jaringan binatang menunjukkan kafein
dalam kopi menguatkan efek karsinogenik
subtansi tertentu. (1)
Sel preneoplasma dapat tumbuh terus pada
kultur jaringan sedangkan sel normal akan
berhenti tumbuh. (4) Sel preneoplasma lebih
tahan terhadap lingkungan yang tidak
mendukung dan kemampuan kloningnya lebih
besar. Kebanyakan sel-sel preneoplasma
beregresi menjadi sel berdiferensiasi normal
tetapi sebagian kecil mengalami perkembangan progresif menjadi sel-sel neoplasma
yang ireversibel. (4,6) Pada akhir fase promosi
terdapat gambaran histologis dan biokomiawi
yang abnormal. (6)
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Fase progresi
Fase ini berlangsung berbulan-bulan. (6)
Pada awal fase ini, sel preneoplasma dalam
stadium metaplasia berkembang progresif
menjadi stadium displasia sebelum menjadi
neoplasma. (4,5) Terjadi ekspansi populasi selsel ini secara spontan dan ireversibel. Sel-sel
menjadi kurang responsif terhadap sistem
imunitas tubuh dan regulasi sel. Pada esofagus
epitel berlapis gepeng berubah atau metaplasia
menjadi epitel selapis torak yang kemudian
berkembang menjadi jaringan dalam keadaan
displasia yang kemudian berkembang menjadi
neoplasma. Pada kolon, polip adalah bentuk
metaplasia. Pada tingkat metaplasia dan
permulaan displasia (ringan sampai sedang)
masih bisa terjadi regresi atau remisi yang
spontan ke tingkat lebih awal yang
frekwensinya makin menurun dengan bertambahnya progresivitas lesi tersebut. Belum
banyak diketahui perubahan yang terjadi dan
faktor-faktor yang mempengaruhinya. Batas
yang pasti perubahan lesi preneoplasma
menjadi neoplasma sulit ditentukan. Pada akhir
fase ini gambaran histologis dan klinis
menunjukkan keganasan. (6)
Penyelidikan terakhir memperlihatkan terjadi
aglutinasi pada permukaan sel kanker sehingga
sel kanker tumbuh terus meskipun terjadi
kontak antar sel. (4) Permukaan sel kanker
mempunyai lebih sedikit neksus (daerah
kontak antar sel). Ini menunjukkan kurangnya
metabolisme dan pertukaran ion-ion antar sel
yang juga menyebabkan sel kanker bertambah
otonom. Hal ini lebih nyata pada keadaan
displasia yang progresif ke arah neoplasma.
Semua perubahan struktur, metabolik dan
kelakuan sel ini terjadi karena mutasi yang
mengenai inti, mitokondria dan membran
endoplasma sel. Kebanyakan sel kanker
mensekresi enzim fibrinolitik yang melarutkan
jaringan ikat di sekitarnya dan faktor
angiogenesis yang menginduksi pembentukan
kapilar darah baru di antara pembuluh darah
yang berdekatan dengan sel kanker untuk
nutrisinya. Pada permukaan sel kanker
terbentuk antigen yang menimbulkan respons
imun selular dan humoral untuk melawan sel
kanker. (4) Antigen permukaan ini sering
ditemukan di jaringan fetus, mempunyai
hubungan dengan derajat diferensiasi sel dan

kekhasannya dipakai sebagai tambahan pada
diagnostik kanker.
Karsinogen kimia
Bahan kimia sudah terdapat dalam
lingkungan manusia dalam waktu lama,
misalnya benzopyrene sudah terdapat dalam
atmosfer dalam kadar rendah sejak api dari
tumbuh-tumbuhan
dipakai
manusia.
Penyelidikan epidemiologis pada manusia
menunjukkan baru terjadi peningkatan jumlah
penderita kanker paru setelah zat ini dihirup
selama 20 tahun atau lebih. (3) Lebih 200 tahun
lalu ditemukan kanker skrotum pada
pembersih cerobong asap karena terpapar
bertahun-tahun dengan jelaga cerobong asap.
Dari penyelidikan pada permulaan abad ke 20
diketahui dalam jelaga cerobong asap dan ter
batu bara terdapat benzopyrene. Benzopyrene
yang dioleskan pada kulit telinga kelinci dan
yang kontak dengan kulit manusia menimbulkan kanker kulit.
Bahan-bahan kimia baru yang karsinogenik
dihasilkan terus menerus dan kadarnya makin
meningkat sebagai akibat perkembangan
industri. Penyelidikan epidemiologis mengalami kesukaran karena akibat suatu bahan
kimia baru diketahui setelah 20 tahun
kemudian atau lebih, kecuali bahan tersebut
sangat tinggi sifat karsinogeniknya dan
paparannya sangat lama. Tidak mungkin untuk
melakukan tes terhadap tiap bahan industri
baru, kecuali yang diproduksi besar-besaran.
Yang dapat dilakukan adalah mengikuti
perkembangan kesehatan beberapa ratus
pekerja yang terpapar suatu bahan kimia baru
selama paling sedikit 25 tahun. Percobaan
binatang dan tes kultur jaringan sangat
membantu untuk menentukan suatu bahan
kimia karsinogenik atau tidak, sebab bila
terbukti karsinogenik, tanpa melihat dosisnya
dapat dianggap bahwa bahan tersebut juga
mungkin karsinogenik untuik manusia. Hanya
diperlukan penyelidikan lebih lanjut untuk
menentukan berapa dosis dan waktu yang
diperlukan untuk menimbulkan kanker pada
manusia. Tetapi hasil negatif tes-tes ini tidak
menjamin bahan tersebut aman untuk manusia.
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Jenis-jenis karsinogen kimia

Polycyclic aromatic hydrocarbon. (1,4,5,8)
Contoh: benzopyrene terdapat dalam asap
rokok, asap mobil dan sebagai produk
pembakaran
tumbuh-tumbuhan
yang
menyebabkan kanker paru; dalam jelaga
cerobong asap dan ter batu bara menyebabkan
kanker kulit. Asap rokok juga menyebabkan
kanker orofarings, esofagus, larings, kandung
kemih, ginjal dan pankreas. (2) Tembakau yang
dikunyah menimbulkan kanker orofarings.
Benzopyrene juga terbentuk bila daging dan
ikan dipanggang dengan arang, diasap atau
digoreng dengan minyak yang sudah dipakai
berkali-kali (1). Benzopyrene juga terdapat
dalam macam-macam makanan. Beberapa
jenis kerang dan ikan dari air yang terpolusi
dapat mengandung benzopyrene, tetapi dari
penelitian epidemiologis dan percobaan
binatang belum ditemukan hubungannya
dengan kanker. Golongan ini di-hidroksilasi
oleh enzim arylhydrocarbon hydroxylase
(dalam limfosit) menjadi karsinogen yang
reaktif.
Aromatic amine. (4,5) Contoh: butter yellow
(dulu dipakai sebagai pewarna mentega
sebelum efek karsinogeniknya pada binatang
diketahui),
insektisida
naphthylamine,
benzidine
dan
3-acetylaminofluorene.
Naphthylamine menyebabkan kanker hati pada
rodentia dan kanker kandung kemih pada
anjing, (4) juga karsinogenik untuk manusia.
Benzidine menyebabkan kanker kandung
kemih pada pekerja industri zat warna.
Golongan ini diaktifkan dulu oleh enzim dalam
sel hati atau ginjal atau sel tubuh lainnya
menjadi karsinogen yang reaktif.
Alkylating. (4,5) Contoh: epoxide, lactone,
nitrogen mustard dan derivatnya. Nitrogen
mustard untuk pengobatan penyakit Hodgkin
menimbulkan kanker lain pada penderita
tersebut misalnya lekemia, kanker kandung
kemih dan limfoma. Termasuk dalam
golongan ini chlorambucil dan busulphan
menimbulkan
lekemia
sedangkan
cyclophosphamide
menimbulkan
kanker
kandung kemih. Untuk lekemia periode
latennya singkat sedangkan kanker solid lebih
lama. Telah lama diketahui golongan ini
bersifat mutagenik berikatan dengan bagian-
bagian molekul DNA menyebabkan kesalahan

pada replikasi DNA.
Nitrosamine. (4,5) Terbentuk dari nitrit dengan
sejumlah amin. Garam nitrit dan nitrat alamiah
terdapat dalam sayur-sayuran, ikan dan daging.
Nitrit digunakan sebagai aditif makanan
(pengawet daging) sejak abad ke 19 dan
peptisida, juga terdapat dalam makanan
sebagai residu obat-obatan. Sumber amin
adalah obat tertentu dan nikotin. Nitrosamine
juga terbentuk pada proses memanggang dan
terdapat dalam asap rokok. (5,8) Nitrosamine
menyebabkan macam-macam kanker pada
spesies binatang percobaan yang berbeda yaitu
kanker hati, ginjal, paru, esofagus, vesika
urinaria, pankreas, trakea, sinus dan saraf tepi.
Di beberapa bagian dunia misalnya India,
mengunyah buah pinang dapat menyebabkan
kanker mulut, farings atau esofagus,
kemungkinan karena nitrosamine dalam buah
pinang. Penyelidikan epidemiologis membuktikan tidak konsistennya hubungan
nitrosamine
dengan
kanker
lambung.
Golongan ini diaktifkan dulu oleh enzim sel
hati atau ginjal atau sel tubuh lainnya menjadi
karsinogen yang reaktif. Berdasar pengetahuan
saat ini nitrosamine pada manusia belum pasti
menimbulkan kanker.
Aflatoxin B1. (1,4,5) Pada permulaan tahun 1960
diisolasi dari jamur Aspergillus flavus yang
tumbuh pada makanan yang disimpan yaitu
kacang tanah, jagung, gandum, kacang polong,
beras, kacang kedelai, buah, daging tertentu,
susu dan keju. Aflatoxin adalah karsinogen hati
pada beberapa spesies binatang. Pada manusia
menyebabkan kanker hati (hepatoma primer),
terdapat bukti bahwa aflatoxin mempunyai
peranan utama untuk terbentuknya kanker hati
di negara tropis sebagai kontaminan dari
makanan karbohidrat, terutama biji-bijian dan
kacang-kacangan.
Aflatoxin juga ditransformasikan dulu oleh enzim sel hati atau ginjal
menjadi karsinogen yang reaktif.
Logam berat. (1,4,5) Senyawa kromium (Cr),
nikel (Ni) dan uranium (Ur) diduga
menyebabkan kanker paru dan sinus
sedangkan
kadmium
(Cd)
diduga
menyebabkan kanker prostat.
Vinylchloride (1,4,5) , pada pekerja pabrik bahan
dasar plastik, polyvinylchloride (PVC) dapat
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menyebabkan kanker hati (angiosarkoma),

kanker paru, otak, darah dan limf. Bungkus
plastik dan tempat makanan plastik yang
menggunakan bahan dasar vinylchloride
menguatirkan
konsumen.
Chloromethylmethylether digunakan secara luas pada
industri kimia sebagai perantara sintesa
organik dapat menyebabkan kanker paru.
Carbontetrachloride pada pekerja plastik dan
pekerja cuci kering menyebabkan kanker hati,
thiourea (zat aditif makanan) pernah digunakan sebelum diketahui sifat karsinogeniknya pada binatang dan urethane (zat
aditif makanan) diduga karsinogenik. (4)
Hidrocarbonchloride
sebagai
peptisida
misalnya DDT, eldrin, dieldrin menyebabkan
kanker hati pada tikus dan lain spesies, pada
manusia belum jelas menyebabkan kanker,
mungkin karena periode latennya belum
diketahui berapa tahun. (1)
Penggunaan pewarna rambut meningkatkan
risiko terkena limfoma non-Hodgkin, penyakit
Hodgkin dan multiple myeloma. (9) Beberapa
jenis kanker diduga disebabkan beberapa
produk seperti deterjen, kosmetik, plastik padat
atau busa, cat, pewarna, semir, pelarut, kertas
dan tinta cetak. (1) Mungkin setelah paparan
lama risiko ini dapat dideteksi di masa yang
akan datang.
Phenacetin diduga penyebab kanker pelvis
renis dan kandung kemih, methoxypsoralen
penyebab kanker kulit, arsen penyebab kanker
kulit dan chlornaphazine penyebab kanker
kandung kemih. (1)
Radiasi
Terdapat 2 macam radiasi yaitu radiasi
ionisasi (misalnya sinar X) dan non-ionisasi
(sinar ultraviolet). (1,10) Keduanya adalah
bagian
dari
spektrum
gelombang
elektromagnetik. Sinar X berasal dari tambang
uranium, kosmik, alat diagnostik penyakit, alat
terapi radiasi, kecelakaan nuklir, bom atom
dan sampah radioaktif. Sinar ultraviolet berasal
dari matahari.
Risiko terkena kanker meningkat pada anak
yang waktu masa fetusnya terkena radiasi
sinar X dari pelvimetri ibunya atau pada anak
yang sel benih ibunya sebelum kehamilan
mengalami mutasi. Peningkatan penggunaan
enersi nuklir dan percobaan senjata nuklir

mempunyai efek jangka panjang dan pendek
radiasi sinar X. Efek jangka pendek menginduksi kanker, sedangkan jangka panjang
menyebabkan kerusakan gen yang diteruskan
kepada generasi mendatang. (10) Dosis kecilpun
dapat menimbulkan kerusakan jaringan, tetapi
berapa besar dosis belum dapat dipastikan.
Risiko menderita lekemia akut adalah yang
pertama diketahui dan sumsum tulang dulu
dianggap organ yang paling sensitif tetapi
sekarang diketahui risiko untuk menderita
tumor ganas padat lebih besar yaitu kanker
kelenjar tiroid, payu dara, paru, kulit, tulang
dan lambung serta organ pencernaan lainnya.
(1,10,11)
Periode laten untuk lekemia adalah
beberapa tahun (2-5 tahun) sedangkan untuk
tumor ganas padat pada umumnya 5-10 tahun
dapat sampai lebih dari 30 tahun. Zat
radioaktif lain misalnya radium, phosphorus
(P32), mesothorium dan thorotrast dapat
menimbulkan lekemia, osteosarkoma, kanker
sinus dan angiosarkoma hati. (1) Radon dari
elemen tanah menimbulkan kanker paru pada
penambang. (6) Batu-batuan rumah banyak
yang mengandung materi radioaktif antara lain
radon, bila kadar gas ini dalam rumah
meningkat 100 kali melebihi batas aman,
kemungkinan menyebabkan kanker paru pada
yang bukan asap rokok sebagai penyebabnya.
Radon merupakan 10-20% penyebab kanker
paru. (6)
Sinar ultraviolet menyebabkan tumor pada
paparan berulang dan dosis tertentu. Jaringan
yang terkena adalah kulit, biasanya kulit pelaut
dan petani, dapat timbul karsinoma sel basal,
karsinoma sel skwamosa atau melanoma
malignum. (1) Lebih dari 75% kanker kulit
adalah karsinoma sel basal muka dan leher.
Pada bibir terutama karsinoma sel skuamosa
dan paling jarang melanoma malignum tetapi
merupakan penyebab kematian utama kanker
kulit. CFC (chlorofluorocarbon) menyebabkan
berkurang tebalnya lapisan ozon di stratosfer
sehingga radiasi ultraviolet matahari lebih
banyak sampai ke permukaan bumi. Orang
yang genetik melaninnya lebih sedikit lebih
tinggi risiko terkena kanker kulit. Penderita
penyakit
genetik
seperti
Xeroderma
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pigmentosa dan albinisme sangat tinggi risiko

terkena kanker kulit.
Terjadinya kanker karena radiasi sinar X dan
ultraviolet menimbulkan sejumlah lesi yang
berbeda pada DNA sel. (6,10,12) Tidak terjadi
kanker bila lesi ini direparasi atau dimodifikasi
oleh proses biologis atau terjadi kematian sel.
(10)
Virus
Banyak kanker pada binatang disebabkan
oleh virus, pada manusia, virus adalah
penyebab kanker tertentu. (6) Virus EbsteinBarr (EBV) suatu virus herpes adalah penyebab
infectious mononucleosis dan limfoma Burkitt
pada anak-anak di Afrika (1,11) , tetapi malaria
yang menimbulkan supresi imunitas tubuh juga
berperan penting. Data epidemiologis dan
deteksi DNA virus Ebstein-Barr dalam sel
limfoma mendukung hubungan virus ini
dengan 2 macam kelainan
tersebut. Di
Amerika Serikat limfoma Burkitt agaknya
tidak mempunyai hubungan dengan virus
Epstein-Barr. Virus ini diduga ko-karsinogen
untuk kanker nasofarings pada orang Cina
dengan peranan faktor genetik lebih besar. (1,11)
Malaria membantu timbulnya kanker ini. Virus
ini juga diduga penyebab kanker timus. (11)
Virus papiloma (HPV) subtipe 6, 8, 16 dan 18
(6,11)
, virus herpes simplex tipe 2 dan virus
cytomegalo (1) berhubungan erat dengan risiko
terkena kanker serviks. Ketiga virus ini juga
diduga penyebab kanker penis. Penularan virus
didapat karena hubungan seksual yang
biasanya terjadi pada orang yang menikah
pada usia muda atau mempunyai banyak
pasangan atau pada populasi sosio-ekonomi
rendah.
Virus hepatitis B (HBV) endemik di
masyarakat tertentu misalnya masyarakat
Taiwan dan negro Afrika Selatan. Virus ini
pada umumnya menyebabkan hepatitis akut
atau infeksi yang asimptomatik yang
menimbulkan kekebalan. Sebanyak 10% dari
penderita berlanjut menjadi kronis, sirosis
hepatis kemudian kanker hati. (11) Virus
menyebabkan
kematian
jaringan
hati,
merangsang pembelahan sel dan membuat
rentan terhadap karsinogen kimia, maka
kanker hati lebih mudah timbul dengan adanya
aflatoxin B1 sebagai ko-karsinogen. Juga

malaria
dan malnutrisi mempermudah
timbulnya kanker ini.
Penularan virus ini
melalui hubungan seksual, plasenta dari ibu ke
anak dan penggunaan jarum suntik intravena
(transfusi darah atau penggunaan obat
terlarang).
Virus lekemia sel T (HTLV-I / human T cell
leukemia virus tipe I) adalah penyebab lekemia
limfosit T orang dewasa di Jepang dan India
Barat, virus ini adalah virus yang langsung
menyebabkan kanker. (11)
HIV (human immuno-deficiency virus) adalah
penyebab sarkoma Kaposi (11) yang termasuk
dalam AIDS (acquired immuno-deficiency
syndrome). Penularan virus ini melalui cara
yang sama seperti virus hepatitis B.
Dalam 10 tahun terakhir hubungan virus
manusia dengan kanker tertentu dipelajari
dengan penyelidikan epidemiologis, klinis dan
biologi molekular. Dari penyelidikan virus
DNA atau virus RNA (retrovirus) yang
membentuk
tumor
ganas
didapatkan
terbentuknya tumor ganas karena onkogen
virus yang masuk ke gen sel penderita yaitu
integrasi sebagian atau seluruh DNA virus atau
DNA copy dari RNA retrovirus ke kromosom
penderita. (11) Dengan cara langsung
pengaktifan onkogen sel penderita atau dengan
cara menghilangkan fungsi gen supresor tumor
penderita sebagai akibat mutasi gen penderita
terbentuk neoplasma. Mekanisme yang
sebenarnya belum diketahui, kemungkinan
mencakup lebih dari hanya integrasi elemen
virus ke kromosom penderita. Tidak
ditemukannya DNA virus dalam materi genetik
sel kanker mungkin karena metode untuk
mendeteksi virus tidak cukup sensitif atau
mungkin karena virus tidak sering mempunyai
hubungan dengan kanker.
Faktor genetik
Tumor masa anak yaitu retinoblastoma
telah lama dipandang sebagai contoh dari
kanker yang diturunkan secara dominan, tetapi
tumor ini dapat juga non-herediter. (1,13) Dari
data keluarga didapatkan kurang dari 50%
keturunan penderita retinoblastoma yang
kemudian akan menderita tumor ini. Mereka
menduga ini termasuk dalam golongan tumor
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yang
non-herediter.
Penyelidikan
lain
menunjukkan
penderita
retinoblastoma
bilateral yang sebelumnya tidak mempunyai
riwayat keturunan kemungkinan menurunkan
penyakit ini mendekati 50% seperti pada
penderita retinoblastoma unilateral yang
mempunyai riwayat keturunan. Sedangkan
kemungkinan mendapat penyakit ini adalah
10-15% pada keturunan dari penderita
retinoblastoma
unilateral
yang
tidak
mempunyai riwayat keturunan. Kemungkinan
mendapat penyakit ini pada keturunan
penderita yang tumornya unilateral atau
bilateral dengan riwayat keturunan sangat
tinggi yaitu 60-70%. Dari data disimpulkan
40% keturunan penderita retinoblastoma
adalah karier gen yang dominan. Dari 40% ini,
95% menderita paling sedikit tumor unilateral
bisa juga bilateral. Sebaliknya penderita yang
tidak membawa gen dominan mempunyai
risiko 1/30.000 untuk menderita tumor
unilateral dan tidak pernah bilateral.
Terdapat dugaan mutasi terjadi pada sel-sel
retina karier gen sehingga terbentuk
retinoblastoma. Juga diduga bentuk herediter
terjadi secara 2 tahap yaitu mutasi sel somatik
(sel retina) yang sedang tumbuh dan mutasi
pada sel benih yang akan diturunkan. Pada
bentuk non-herediter terjadi 2 tahap mutasi
yang ke 2 nya terjadi dalam sel somatik untuk
menjadi sel kanker tetapi fenomena ini sangat
jarang (1/30.000). Maka pada bentuk herediter
retinoblastoma terjadi lebih awal dari bentuk
non-herediter karena hanya diperlukan 1 tahap
yang terjadi post-zygotik.
Tumor lain yang mempunyai bentuk herediter
dan non-herediter adalah tumor Wilm (ginjal),
pheochromocytoma (ginjal) (13), kanker kolon,
karsinoma sel basal dan lekemia. (1) Bentuk
herediter tumor Wilm dan pheochromocytoma
timbul pada masa anak. Kanker kolon dan
karsinoma sel basal yang timbul pada usia
muda adalah bentuk herediter.
Faktor keturunan pada kanker didasarkan pada
pengamatan klinis dan laboratoris (terdapatnya
kelainan kromosom yang khas untuk kanker
tertentu).(1) Pada retinoblastoma hilangnya
sebagian kromosom 13, pada tumor Wilm
hilangnya sebagian kromosom 11 dan pada
lekemia kelainan trisomy 21.
Tumor pada masa anak-anak dapat tidak

menjadi kanker dengan bertambahnya usia
karena sel genetis yang berpotensi menjadi
kanker dengan bertambahnya umur mengalami
diferensiasi dan menghilang.
Faktor psikogenik (14)
Faktor ini mencakup 2 bagian yaitu faktor
kepribadian dan psikososial.
Penyelidikan-penyelidikan terhadap hubungan
kepribadian dengan kanker mendapatkan hasil
yang berlawanan, misalnya tes psikologi pada
wanita yang akan dibiopsi mammanya
mendapatkan emosi yang tertekan pada
penderita kanker mamma tetapi penyelidikan
lain tidak membuktikan hubungan stres dengan
kanker mamma.
Hubungan antara kejadian yang menekan
dalam kehidupan seseorang dengan penyakit
(faktor psikososial) telah lama diselidiki.
Kejadian yang menekan ini misalnya
perceraian,
keadaan
menganggur
dan
kehilangan orang yang dicintai atau rumah.
Penyelidikan mendapatkan bahwa pada
penderita kanker lebih banyak yang mengalami
kehilangan objek yang dicintai dari orang yang
tidak menderita kanker. Penyelidikan lain
mendapatkan makin banyak stres makin
banyak kemungkinan seseorang sakit karena
penurunan imunitas tubuh. Terdapat teori yang
mengatakan bukan kejadian dalam hidup
seseorang yang merupakan predisposisi untuk
kanker tetapi
mekanisme pertahanannya
(penyangkalan, represi atau proyeksi bisa
sampai putus asa), tergantung kemampuan
seseorang untuk mengatasi pengalaman yang
menekan tersebut. Telah lama dikenal postulat
tentang sistem simpatis-meduloadrenal dan
poros pituitari-korteks adrenal. Emosi oleh
korteks serebri diteruskan sebagai impuls ke
hipotalamus, dari sini merangsang sistem
simpatis dan kelenjar pituitari. Sistem simpatis
merangsang
pengeluaran
katekolamin
(adrenalin dan noradrenalin) dari medula
adrenal dan kelenjar pituitari melepaskan
hormon yang berefek kepada korteks adrenal
dan kelenjar tiroid. Stres menyebabkan
perubahan aktifitas sistem saraf dan endokrin,
perubahan
pada
sistem
endokrin
mengakibatkan kenaikan sekresi hormon
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kortikosteroid dari korteks adrenal yang

menyebabkan involusi kelenjar timus, ini
menurunkan jumlah limfosit T yang
menyebabkan penurunan imunitas tubuh
sehingga seseorang menjadi rentan terhadap
kanker.
Pencegahan
Penyakit manusia yang utama kebanyakan
dikontrol tidak secara agresif tetapi lebih
secara aplikasi penemuan pengetahuan dasar
yang
mendukung
strategi
kesehatan
masyarakat. (6) Juga untuk kanker yang paling
baik dan efektif adalah pencegahan.
Pencegahan primer
Kemajuan
dalam
bidang
biologi
molekular telah dapat mengetahui pentingnya
suatu materi genetik yang hilang pada fase
inisiasi
banyak
kanker
misalnya
retinoblastoma dan kanker kolon yang
herediter. Kelainan ini telah dapat dikoreksi in
vitro dengan cara pemindahan gen via vektor
retrovirus. Penyempurnaan teknik ini untuk
penggunaan in vivo mungkin berperanan
penting untuk mengurangi insidens kanker di
masa yang akan datang. Penting diselidiki
apakah terjadi perubahan mendasar pada gen
tertentu orang yang berisiko tinggi untuk
menderita kanker dan diselidiki gen-gen
spesifik yang berhubungan erat dengan
macam-macam kanker. Bila pada gen seorang
individu terdapat perubahan mendasar yang
meningkatkan risiko untuk menderita kanker
tertentu, dilakukan skrining dan deteksi awal
juga diberikan nasihat untuk menaikkan
kondisi kesehatannya. Pada individu yang
menderita poliposis kolon familial dilakukan
colonoscopy dan colectomy aksis.
Menghindari kontak dengan karsinogen kimia,
radiasi atau virus termasuk dalam pencegahan
primer yaitu mencegah terjadinya sel
terinisiasi. Strategi pencegahan primer yang
umum dan telah lama diketahui misalnya tidak
merokok untuk pencegahan kanker paru dan
penggunaan tabir surya untuk pencegahan
kanker kulit. Juga termasuk dalam pencegahan
primer adalah menghindari faktor-faktor yang
dapat menginduksi replikasi DNA.
Saat ini sedang aktif diselidiki efek antiinisiator (penghambat inisiasi) misalnya
vitamin E, C, beta-karoten, selenium, ekstrak
kacang kedelai (penghambat protease) dan
anti-oksidan lain. Belum ditemukan cara
efektif untuk mengatasi kualitas udara buruk
dalam rumah. (6)
Pencegahan sekunder
Pencegahan ini ditujukan terhadap sel
terinisiasi dengan cara menghindari paparan
dengan promotor misalnya alkohol, hormon,
lemak, sakarin dan siklamat. Memperbanyak
substansi yang mengurangi paparan dengan
promotor misalnya serat dalam makanan dan
substansi yang memperbaiki lingkungan
jaringan sehingga proliferasi sel terinisiasi
ditekan atau diferensiasi sel ditingkatkan
misalnya konsumsi adekuat vitamin E, C,
beta-karoten dan vitamin A.
Pencegahan tersier
Paling sering digunakan operasi atau
ablasi untuk preneoplasma. (6) Tindakan ini
tidak selalu mudah dilakukan dan lesi sering
timbul kembali. Saat ini sedang diselidiki
intervensi aktif yang memperbaiki lingkungan
seperti zat anti-proliferatif atau zat diferensiasi
untuk mengembalikan keadaan preneoplasma
ke fase sebelumnya atau menghambat
progresifitas preneoplasma menjadi neoplasma
misalnya lekoplakia mulut dan displasia
serviks dengan retinoid atau beta-karoten (6)
atau asam folat. (1) Keganasan tertentu in vitro
dan pada binatang percobaan yang sensitif
terhadap zat diferensiasi misalnya sel
neuroblastoma in vitro berdiferensiasi sampai
matang dengan adanya asam retinoat. Belum
dilakukan percobaan apakah hal ini juga
berlaku in vivo. Beberapa tahun terakhir ini
diketahui tulang rawan ikan hiu dapat
menghambat angiogenesis. Juga ditemukan zat
dalam jamur maitake yang dapat menghambat
progresifitas
neoplasma
dengan
cara
memperkuat sistem imunitas tubuh.
Faktor psikogenik mempengaruhi tiap fase
karsinogenesis karena mempunyai hubungan
dengan imunitas tubuh.
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KESIMPULAN
Dari penyelidikan epidemiologis dan
laboratoris (percobaan binatang dan kultur
jaringan) didapatkan bahwa karsinogen kimia
merupakan penyebab kanker yang utama dan
paling banyak diselidiki. Penyebab kanker
yang lain adalah radiasi, virus, faktor genetik
dan faktor psikogenik. Selain ini terdapat zat
yang disebut promotor yang membantu
terjadinya kanker.
Dengan memperhatikan macam-macam penyebab kanker dan promotor, kemungkinan
menderita kanker dapat dikurangi seminimal
mungkin. Di masa akan datang kemungkinan
kemajuan ilmu biologi molekular ikut
berperanan menurunkan insidens kanker.
DAFTAR PUSTAKA
1.
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5.
Hammond EC. The epidemiological approach

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JL, Hart LK, editors. Cancer pathophysiology,
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Chyou PH. A prospective study of the
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Rauth AM. Radiation carcinogenesis. In:
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Archer MC. Chemical carcinogenesis. In:
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Benchimol S. Viruses and cancer. In: Tannock
JF, Hill RP, editors. The basic science of
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6. Ryser HJP. Chemical carsinogenesis. In: Kruse
LC, Reese JL, Hart LK, editors. Cancer
pathophysiology, etiology and management. 4th
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7. Deininger MWN. Selective induction of
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421-5.
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editor.
Clinical oncology. 1st ed. Texas:
Appleton & Lange; 1993. p. 12-6.
9. Headley DB. Premorbid psychological factors
of cancer. In: Kruse LC, Reese JL, Hart LK,
editors. Cancer pathophysiology, etiology and
management. 4th ed. St Louis: The C.V. Mosby
Co.; 1975. p. 63-72.
10. Meyskens Jr FL. Cancer prevention. In: Weiss
GR, editor. Clinical oncology. 1st ed. Texas:
Appleton & Lange; 1993. p. 41-7.
11. Knudson Jr AG. Hereditary and human cancer.
In: Finney MI, editor. Cancer risk. assessing
and reducing the dangers in our society. 1st ed.
Colorado: Westview Press, Inc.; 1982. p. 5761.
12. Gibbons JH. Factors associated with cancer. In:
Finney MI, editor. Cancer risk. Assessing and
reducing the dangers in our society 1st ed.
Colorado: Westview Press, Inc.; 1982. p. 3-9,
78-85.
13. Giovannucci E. Calcium and fructose intake
in relation to risk of prostate cancer. Cancer
Research 1998;58 : 443-6.
14. Zahm SH. Use of hair coloring products and
the risk of lymphoma, multiple myeloma and
chronic lymphocytic leukemia. Am J Publ
Health1992:82:990-6
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Research Article
Polychlorinated Biphenyls and Risk of Testicular Germ Cell Tumors

1
Katherine A. McGlynn, Sabah M. Quraishi, Barry I. Graubard, Jean-Philippe Weber,

3
4
Mark V. Rubertone, and Ralph L. Erickson
1
Division of Cancer Epidemiology and Genetics, National Cancer Institute, NIH, Department of Health and Human Services,
Bethesda, Maryland; 2Toxicology Centre, Institut National de Sante Publique du Quebec, Quebec City, Quebec, Canada;
and 3U.S. Army Center for Health Promotion and Preventive Medicine; 4Global Emerging Infections Surveillance and
Response System, Walter Reed Army Institute of Research, Silver Spring, Maryland
Abstract
Exposure to endocrine-disrupting chemicals, such as polychlorinated biphenyls (PCB), may alter hormonal balance and
thereby increase risk of testicular germ cell tumors (TGCT). To
study the relationship of PCBs to TGCT, prediagnostic serum
samples from 736 cases and 913 controls in the Servicemens
Testicular Tumor Environmental and Endocrine Determinants
study were analyzed. Adjusted odds ratios and 95% confidence
intervals were estimated using logistic regression. PCB levels
were examined in association with all TGCT and, separately,
with each histologic type (seminoma and nonseminoma).
Risks associated with seven functional groupings of PCBs,
as well as sum of PCBs, were also examined. There were
significantly decreased risks of TGCT in association with
eight PCBs (PCB-118, PCB-138, PCB-153, PCB-156, PCB-163,
PCB-170, PCB-180, and PCB-187) and no association with
the remaining three (PCB-99, PCB-101, and PCB-183). The
same eight congeners were significantly associated with decreased risk of nonseminoma, whereas five (PCB-138, PCB153, PCB-156, PCB-163, and PCB-170) were associated with
decreased risk of seminoma. All functional groupings of PCBs
were also associated with decreased risk of TGCT and of
nonseminoma, whereas six of the seven functional groups
were associated with decreased risk of seminoma. Sum of
PCBs was significantly associated with decreased risk of TGCT
(P trend = 0.006), nonseminoma (P trend = 0.007), and seminoma
(P trend = 0.05). Overall, these data do not support the
hypothesis that PCB exposure increases the risk of TGCT.
[Cancer Res 2009;69(5):19019]
Introduction
The incidence of testicular germ cell tumors (TGCT) has been
increasing in the United States and in other developed countries
for at least 5 decades (1). Reasons for the increase are not clear, as
the etiology of TGCT is poorly understood. The only well-described
risk factors are cryptorchism, prior history of TGCT, family history
of TGCT, and increased adult stature (2). The association of TGCT
with a congenital anomaly regulated by androgen levels, as well as
the similarity between testicular carcinoma in situ and primordial
germ cells, has suggested that TGCT risk may be determined
Note: Supplementary data for this article are available at Cancer Research Online
(http://cancerres.aacrjournals.org/).
Requests for reprints: Katherine A. McGlynn, Hormonal and Reproductive
Epidemiology Branch, Division of Cancer Epidemiology and Genetics, National Cancer
Institute, NIH, Department of Health and Human Services, EPS-Suite 550, 6120
Executive Boulevard, Rockville, MD 20852-7234. Phone: 301-435-4918; Fax: 301-4020916; E-mail: mcglynnk@mail.nih.gov.
I2009 American Association for Cancer Research.
doi:10.1158/0008-5472.CAN-08-3935
www.aacrjournals.org
very early in life and may be affected by hormone levels (37).

The ability of exogenous exposures, such as synthetic hormonal
drugs, phytoestrogens, and persistent organochlorine compounds,
to mimic the effects of endogenous hormones has led to speculation that these exposures might also be related to the development of TGCT and the other male reproductive disorders
(cryptorchism, hypospadias, and impaired spermatogenesis) collectively known as the testicular dysgenesis syndrome (8, 9).
Recently, we reported that p,p-dichlorodiphenyldichloroethylene
(p,p-DDE), the most persistent metabolite of 1,1,1-trichloro-2,2bis(p-chlorophenyl)ethane; dichlorodiphenyltrichloroethane, was
significantly associated with increased risk of TGCT (10). Like
p,p-DDE, other organochlorine compounds share the ability to act
as endocrine disruptors. Whereas p,p-DDE has been shown to have
antiandrogenic properties (11), various polychlorinated biphenyl
(PCB) congeners are known to have estrogenic, antiestrogenic,
androgenic, and antiandrogenic properties (12). The effect of PCBs
on TGCT risk could then be similar to that of p,p-DDE or could
be dissimilar depending on the mix of PCB congeners present. One
prior TCGT case-control study, conducted in Sweden, found no
association between TGCT and total PCB level (13). The study,
however, did find that mothers of the case men had significantly
higher total PCB levels than did mothers of the control men (13).
To pursue this lead and to examine the relationship between TGCT
and specific PCBs, a study was conducted among members of the
U.S. Armed Forces.

Study population. The source population for the current study is
>4 million U.S. military personnel whose blood samples are stored at 30jC
in the Department of Defense Serum Repository (DoDSR) in Silver Spring,
Maryland (14). In the late 1980s, the DoDSR began storing surplus sera from
HIV testing of military personnel. The Repository now includes >40 million
samples from active-duty and reserve personnel, with f2.3 million samples
added each year. Blood samples are drawn at the time of service entry
and, on average, are collected every 2 y thereafter. The earliest blood
sample available from each individual was selected for the current study.
The blood samples included in the current study were donated between
1987 and 2002.
Case identification was accomplished by linking the DoDSR computerized database, using a person-specific ID, to the Defense Medical
Surveillance System to determine which of the serum donors had developed
testicular cancer after donation. The histology of all reported testicular
cancers was determined by examination of the original pathology reports
or by review (6.5%) of the pathology slides. As the study was focused on
TGCTs, the histologies were limited to classic seminoma or nonseminoma
(embryonal carcinoma, yolk sac carcinoma, choriocarcinoma, teratoma,
and mixed germ cell tumor). The database linkage identified 961 case men
who seemed to meet the study criteria. Of these men, 76 could not be
traced, 27 had died, 3 were deployed, and 2 were deemed ineligible, leaving
853 possible participants. Of these men, 22 were in the process of being
1901
Cancer Res 2009; 69: (5). March 1, 2009
Cancer Research
contacted when the study closed. Thus, of the 831 men contacted, 754 (91%)
agreed to participate. In the instances where the potential case participant
was deceased (n = 27), the study attempted to obtain proxy information
from the mans mother. Thirteen proxy questionnaires were completed. The
TGCT cases were diagnosed between 1988 and 2003.
Men with a sample in the DoDSR who had not subsequently developed
testicular cancer were eligible to participate as controls. The study was
designed as a pair-matched case-control study, although additional controls
were initially identified due to the transient nature of the military
population. From the list of all possible controls for each case, four
individuals who matched on birth date (within 1 y), race/ethnicity (white,
black, and other), and date of available serum sample (within 30 d) were
chosen at random as the control set. Among the controls, 2,579 were
evaluated for inclusion. Of these men, 385 could not be traced, 18 had died,
64 were deployed, 2 were deemed ineligible, and 928 could not be contacted
within 30 d. Of the remaining 1,182 men, 32 were in the process of being
contacted when the study closed. Thus, of the 1,150 men contacted, 928
(81%) agreed to participate. Among the 754 cases and 928 controls enrolled,
720 were matched case-control pairs. The volume of serum in the samples
of 18 cases and 15 controls was insufficient to conduct PCB analysis; thus,
the final analysis set included 736 cases and 913 controls.
Study participation included completion of the study questionnaire,

donation of a buccal cell mouthwash sample, permission to use a DoDSR
serum specimen, and written informed consent to participate. In addition,
each participant was asked for permission to contact his mother to
enroll her in the study. For 30 d, every attempt was made to enroll the
first control man in the set via tracing attempts, multiple letters, and
telephone calls. If the potential control could not be contacted, the
enrollment process began anew with the next man in the set. All participants
were enrolled between April 2002 and January 2005. The study was approved
by Institutional Review Boards of the National Cancer Institute (Bethesda,
MD) and the Walter Reed Army Institute for Research (Silver Spring, MD).
Questionnaire data. Each participant was administered a computerassisted telephone interview composed of nine modules. Cases were asked
questions in reference to a date 1 y before their TGCT diagnosis (referred to
as the reference date). Control participants were assigned the same
reference date as their matched case. For the current analysis, participants
were asked to report whether they had a personal history of cryptorchism, a
family history of testicular cancer in first and second relatives, and their
height and weight as of the reference date.
Laboratory methods. Levels of 15 PCB congeners were analyzed by the
Human Toxicology Laboratory of the Institut National de Sante Publique du
Table 1. Selected characteristics of cases and controls in the STEED study

Controls (n = 913)
n (%)
Reference age (y)
1820
2125
2630
3135
3640
4145
Race/ethnicity
White
Black
Other
History of cryptorchism
No
Yes
c
Family history of testicular cancer
No
Yes
b
BMI
<18.5
18.524.9
25.029.9
z30.0
Adult height (in)
<68
6870
7172
z73
DDE (ng/g lipid)
Q1 (V157)
Q2 (158250)
Q3 (251390)
Q4 (>390)
All TGCT (n = 736)

n (%)
P*
Seminoma (n = 313)
n (%)
0.94
67
306
260
159
95
26
(7.3)
(33.5)
(28.5)
(17.4)
(10.4)
(2.8)
64
238
205
132
75
22
(8.7)
(32.3)
(27.9)
(17.9)
(10.2)
(3.0)
623 (84.6)
22 (3.0)
91 (12.4)
897 (98.2)
16 (1.8)
697 (94.7)
39 (5.3)
899 (98.5)
14 (1.5)
705 (95.8)
31 (4.2)
11
404
451
47
(1.2)
(44.2)
(49.4)
(5.1)
9
321
359
45
(1.2)
(43.6)
(48.8)
(6.1)
257
253
237
168
(28.09)
(27.65)
(25.90)
(18.36)
151
217
193
178
(20.43)
(29.36)
(26.12)
(24.09)
238
230
220
224
(26.10)
(25.22)
(24.12)
(24.56)
186
167
146
236
(25.31)
(22.72)
(19.86)
(32.11)
n (%)
<0.0001
12
57
106
74
49
15
(3.8)
(18.2)
(33.9)
(23.6)
(15.7)
(4.8)
0.47
780 (85.4)
34 (3.7)
99 (10.8)
P*
Nonseminoma (n = 422)
<0.0001
52
180
99
58
26
7
(12.3)
(42.7)
(23.5)
(13.7)
(6.2)
(1.7)
0.10
253 (80.8)
12 (3.8)
48 (15.3)
<0.0001
0.37
370 (87.7)
10 (2.4)
42 (10.0)
0.07
302 (96.5)
11 (3.5)
0.0009
<0.0001
394 (93.4)
28 (6.6)
0.0004
297 (94.9)
16 (5.1)
0.86
0.02
407 (96.4)
15 (3.6)
0.24
4
136
147
26
(1.3)
(43.5)
(47.0)
(8.3)
60
96
84
73
(19.17)
(30.67)
(26.84)
(23.32)
59
68
57
128
(18.91)
(21.79)
(18.27)
(41.03)
0.0009
0.93
4
185
212
19
(0.9)
(43.8)
(50.2)
(4.5)
91
121
109
104
(21.41)
(28.47)
(25.65)
(24.47)
127
98
89
108
(30.09)
(23.22)
(21.09)
(25.59)
0.01
0.005
P*
0.02
<0.0001
0.33
*P value of m2 test.
cFamily history in first- and second-degree relatives.
bBMI = height/weight2 (kg/m2).
Cancer Res 2009; 69: (5). March 1, 2009
1902
PCBs and Testicular Germ Cell Tumors
Table 2. Pearson correlation coefficients of PCBs among controls in the STEED study
PCB
PCB-99
PCB-101
PCB-118
PCB-138
PCB-153
PCB-156
PCB-163
PCB-170
PCB-180
PCB-183
PCB-187
Sum of PCBs
Wolff 1B
Wolff 2A
Wolff 2B
Wolff 3
CYP inducers
Phenobarbital inducers
Mixed function oxidase inducers
Wolff groups
101 118 138 153 156 163 170 180 183 187 Sum
1B
2A
2B
CYP
PB
MFO
0.26
0.45
0.67
0.66
0.69
0.77
0.62
0.73
0.75
0.80
0.77
0.86
0.83
0.70
0.32
0.92
0.93
0.91
0.92
0.92
0.76
0.72
0.72
0.69
0.93
0.69
0.58
0.21
0.73
0.94
0.97
0.92
0.94
0.94
0.90
0.87
0.86
0.98
0.76
0.89
0.70
0.27
0.76
0.93
0.97
0.86
0.92
0.87
0.88
0.92
0.87
0.98
0.80
0.88
0.96
0.69
0.38
0.83
0.94
0.97
0.88
0.94
0.87
0.87
0.89
0.87
1.00
0.86
0.93
0.96
0.98
0.69
0.39
0.80
0.93
0.97
0.87
0.94
0.87
0.87
0.90
0.86
0.99
0.86
0.91
0.95
0.99
1.00
0.66
0.27
0.85
0.96
0.97
0.95
0.96
0.88
0.83
0.82
0.80
0.98
0.75
0.97
0.97
0.94
0.97
0.96
0.71
0.45
0.73
0.23
0.81
0.63
0.24
0.77
0.96
0.59
0.15
0.70
0.90
0.91
0.59
0.21
0.75
0.91
0.96
0.95
0.38
0.18
0.57
0.78
0.87
0.83
0.87
Quebec. The PCBs examined, by IUPAC number, were PCB-28, PCB-52,

PCB-99, PCB-101, PCB-105, PCB-118, PCB-128, PCB-138, PCB-153, PCB-156,
PCB-163, PCB-170, PCB-180, PCB-183, and PCB-187. The plasma samples,
enriched with isotopically labeled internal standards, were denatured using
formic acid. Analytes were then automatically extracted from the matrix by
solid-phase extraction. Extracts were automatically cleaned on a florisil
column and analyzed by gas chromatography-mass spectrometry. Detection
of ions generated from negative chemical isolation was accomplished in the
selected ion monitoring mode. Evaluation of concentrations was done by
considering recoveries of the labeled internal standards. Linear calibrations
extended up to 10 Ag/L for most analytes. Higher concentrations were
determined after appropriate dilutions. The mean detection limits
approximated to 0.005 Ag/L. All control samples were run in the same
batch, on the same day, as their matched case sample. Average within-day
variability ranged from 2% to 5%. Average recoveries were 80%. The PCB
limits of detection, median levels, and coefficients of variation, which
include both within- and between-batch variability determined from
replicate QC samples, are shown in Supplementary Table S1. Accuracy of
results was validated through successful participation in two external
quality assessment schemes: the German External Quality Assurance
Scheme (Erlangen University) and the AMAP Ring Test (Quebec).
To adjust all measurements for lipid levels, samples were analyzed for
triglycerides, free and total cholesterol, and phospholipids. The measurements were made with enzyme bioassays using kits produced by Randox
Laboratories. Measurements of PCBs and lipid levels were obtained for 739
cases and 915 controls.
In addition to analyzing each congener separately, the relationships
of TGCT to previously suggested groupings of congeners were examined.
The groupings used were as follows: Wolff groupings (15): Wolff group 1B
(PCB-101 and PCB-187), Wolff group 2A (PCB-118 and PCB-156), Wolff
group 2B (PCB-138 and PCB-170), and Wolff group 3 (PCB-99, PCB-153,
PCB-180, and PCB-183); phenobarbitol inducers (PCB-99, PCB-101, PCB153, PCB-163, PCB-180, and PCB-183; ref. 16); mixed function oxidase
inducers (PCB-118, PCB-138, PCB-156, and PCB-170; ref. 16); and UDP-GT,
CYP1A, and CYP2B inducers (PCB-99, PCB-101, PCB-118, PCB-153, PCB-156,
PCB-180, PCB-183, and PCB-187; ref. 17). The PCBs in each of the groupings
were calculated by summing over the individual results.
Statistical analysis. To conduct logistic regression analyses, lipidadjusted PCB levels were categorized into quartiles based on the levels in
Inducers
0.34
0.21
0.54
0.74
0.86
0.78
0.84
0.96
0.48
0.23
0.62
0.82
0.88
0.72
0.80
0.81
0.84
0.41
0.20
0.56
0.75
0.85
0.71
0.82
0.86
0.91
0.85
0.67
0.34
0.82
0.95
0.98
0.90
0.95
0.88
0.87
0.86
0.86
controls. PCB levels that fell below or were equal to the limit of detection
were imputed as the midpoint of the limit and were included in the first
quartile for regression analysis. Odds ratios (OR) and 95% confidence
intervals (95% CI) were calculated to estimate the association of each PCB
with risk of TGCT and separately with risk of seminoma and nonseminoma.
One case was excluded from histology-specific analyses because tumor
histology was unavailable.
Given the matched case-control design, risk estimates adjusting for
confounders were first generated using conditional logistic regression,
restricting the analysis to only the matched sets. Modeling using
unconditional logistic regression was subsequently performed using the
data from all participants. As the latter involved breaking the match, risk
estimates derived from the unconditional logistic regression models were
adjusted for the three matching factors: age at reference date, race/
ethnicity, and date of serum sample collection. Both logistic regression
models were adjusted for cryptorchism, family history of testicular cancer,
age at serum draw, adult stature, and body mass index (BMI), as BMI is
associated with plasma PCB levels (18). As previous research in the STEED
population had identified p,p-DDE as a risk factor, serum p,p-DDE level
was also included in the models (10). Tests for trend in risk were computed
using scored variables for PCB levels, based on the median levels of each
quartile, to evaluate possible dose-response relationships. As results using
conditional and unconditional logistic regression were similar, only those
using the latter approach are presented.
All tests were two sided, with P < 0.05 defined as the level of statistical
significance.
Statistical analyses were conducted using Statistical Analysis System
Release 9.1 (SAS Institute, Inc.).
Results
As shown in Table 1, 913 controls and 736 cases (313 seminoma, 422 nonseminoma, and 1 histology unavailable) were included in the analysis. Cases and controls were matched on age
and ethnicity, resulting in no significant differences in the distributions of these variables. The seminoma cases were somewhat
older than the controls, whereas the nonseminoma cases were
somewhat younger. The cases, particularly the nonseminoma cases,
1903
Cancer Res 2009; 69: (5). March 1, 2009
Cancer Research
Table 3. Adjusted relative risk of TGCTs by quartile of lipid-adjusted levels of serum organochlorines in the STEED study
ng/g lipid
PCB-99
Q1
Q2
Q3
Q4
P trend
PCB-101
Q1
Q2
Q3
Q4
P trend
PCB-118
Q1
Q2
Q3
Q4
P trend
PCB-138
Q1
Q2
Q3
Q4
P trend
PCB-153
Q1
Q2
Q3
Q4
P trend
PCB-156
Q1
Q2
Q3
Q4
P trend
PCB-163
Q1
Q2
Q3
Q4
P trend
PCB-170
Q1
Q2
Q3
Q4
P trend
PCB-180
Q1
Q2
Q3
Q4
P trend
Controls
All TGCT
Seminoma
Nonseminoma
OR* (95% CI)
OR* (95% CI)
OR* (95% CI)
<9.9
9.912.5
12.618.3
>18.3
590
109
106
107
496
68
90
81
1.00
0.72 (0.521.02)
0.89 (0.641.24)
0.80 (0.571.13)
0.14
199
33
40
41
1.00
0.78 (0.501.22)
0.86 (0.561.34)
0.80 (0.511.25)
0.26
297
35
49
40
1.00
0.68 (0.441.04)
0.91 (0.611.35)
0.76 (0.501.17)
0.16
<4.7
4.76.1
6.28.9
>8.9
570
114
114
114
469
82
93
90
1.00
0.93 (0.671.27)
1.06 (0.781.44)
1.01 (0.741.38)
0.92
200
33
39
40
1.00
0.82 (0.531.27)
1.06 (0.701.61)
1.12 (0.741.70)
0.64
268
49
54
50
1.00
0.99 (0.681.45)
1.12 (0.771.62)
0.91 (0.621.33)
0.80
<7.2
7.210.5
10.615.6
>15.6
265
216
216
216
266
171
151
148
1.00
0.71 (0.530.94)
0.60 (0.450.81)
0.55 (0.400.76)
0.0007
82
80
76
75
1.00
0.97 (0.661.44)
0.86 (0.571.28)
0.72 (0.471.12)
0.10
184
91
74
73
1.00
0.59 (0.420.82)
0.47 (0.330.68)
0.45 (0.310.66)
0.0001
<15.6
15.624.5
24.637.7
>37.7
236
227
225
225
242
168
162
164
1.00
0.65 (0.480.88)
0.54 (0.390.75)
0.46 (0.320.66)
0.0001
73
68
88
84
1.00
0.78 (0.511.20)
0.78 (0.501.22)
0.52 (0.310.86)
0.01
169
100
74
79
1.00
0.61 (0.430.86)
0.41 (0.280.61)
0.42 (0.270.65)
0.0002
<23.4
23.437.2
37.356.3
>56.3
231
227
229
226
243
158
166
169
1.00
0.61 (0.450.82)
0.53 (0.380.73)
0.45 (0.310.66)
0.0003
74
61
85
93
1.00
0.71 (0.461.09)
0.69 (0.441.09)
0.52 (0.310.87)
0.02
169
97
81
75
1.00
0.58 (0.410.82)
0.44 (0.300.65)
0.40 (0.260.63)
0.0002
<5.3
5.36.9
7.010.0
>10.0
477
145
149
142
422
98
120
96
1.00
0.66 (0.480.90)
0.77 (0.561.06)
0.57 (0.400.81)
0.002
148
44
69
52
1.00
0.68 (0.451.05)
0.93 (0.621.40)
0.54 (0.340.86)
0.02
274
54
50
44
1.00
0.63 (0.430.92)
0.63 (0.420.93)
0.58 (0.370.91)
0.006
<5.9
5.98.1
8.211.5
>11.5
397
172
172
172
367
128
110
131
1.00
0.70 (0.520.93)
0.55 (0.400.76)
0.59 (0.420.83)
0.001
126
56
59
72
1.00
0.73 (0.481.09)
0.64 (0.420.98)
0.58 (0.370.92)
0.02
241
72
50
59
1.00
0.69 (0.490.98)
0.46 (0.310.69)
0.57 (0.370.86)
0.002
<6.5
6.59.7
9.814.5
>14.5
335
192
193
193
311
145
136
144
1.00
0.73 (0.550.98)
0.61 (0.440.84)
0.56 (0.390.80)
0.002
101
61
71
80
1.00
0.74 (0.491.12)
0.68 (0.441.04)
0.56 (0.350.91)
0.03
210
84
65
63
1.00
0.74 (0.531.05)
0.56 (0.380.82)
0.55 (0.360.85)
0.005
<15.8
15.825.9
26.041.8
>41.8
234
227
226
226
222
177
176
161
1.00
0.83 (0.621.12)
0.68 (0.490.95)
0.56 (0.380.82)
0.003
65
65
90
93
1.00
0.97 (0.631.48)
0.86 (0.551.37)
0.67 (0.391.13)
0.08
157
112
85
68
1.00
0.80 (0.571.12)
0.59 (0.400.87)
0.51 (0.320.81)
0.004
(Continued on the following page)
Cancer Res 2009; 69: (5). March 1, 2009
1904
Table 3. Adjusted relative risk of TGCTs by quartile of lipid-adjusted levels of serum organochlorines in the STEED
study (Contd)
ng/g lipid
PCB-183
Q1
Q2
Q3
Q4
P trend
PCB-187
Q1
Q2
Q3
Q4
P trend
Controls
All TGCT
Seminoma
Nonseminoma
OR* (95% CI)
OR* (95% CI)
OR* (95% CI)
<4.2
4.25.1
5.26.6
>6.6
689
74
75
75
573
44
54
65
1.00
0.62 (0.410.93)
0.75 (0.501.12)
0.86 (0.581.29)
0.13
219
24
36
34
1.00
0.65 (0.381.09)
0.91 (0.561.47)
0.77 (0.461.29)
0.25
353
20
18
31
1.00
0.56 (0.320.96)
0.55 (0.310.97)
0.92 (0.561.52)
0.14
<5.8
5.88.0
8.111.6
>11.6
386
177
175
175
350
133
120
133
1.00
0.70 (0.520.94)
0.58 (0.420.81)
0.60 (0.420.86)
0.004
111
59
62
81
1.00
0.81 (0.541.21)
0.71 (0.461.10)
0.75 (0.471.20)
0.27
239
74
57
52
1.00
0.65 (0.450.92)
0.49 (0.330.73)
0.48 (0.310.75)
0.0004
*Adjusted for matching variables, serum DDE level, age at serum draw, BMI, and height.
were significantly more likely than the controls to have a history

of cryptorchism (OR, 3.3; 95% CI, 1.85.9; P < 0.0001). The cases
were also significantly more likely to have a family history
of testicular cancer (OR, 2.9; 95% CI, 1.55.5; P = 0.0009), to be
taller (P = 0.0009), and to have higher serum levels of p,p-DDE
(P = 0.005) than the controls. No difference (P = 0.86) in BMI was
evident, however.
Before examining the relationship between PCBs and TGCT, two
preliminary analyses were conducted: an examination of the length
of the interval between serum collection and diagnosis, and a
comparison of mean total lipid levels among cases and controls. The
mean and median intervals between serum collection and diagnosis/
reference date were 3.9 and 3.3 years, respectively. A comparison of
the intervals between the cases and controls found no significant
difference (P = 0.89). Similarly, a comparison of the mean lipid levels
between the cases and controls found no significant difference
(P = 0.11; data not shown). As persons were exposed to mixtures
of PCBs, rather than to single PCB congeners, the correlations
between the congeners and the groupings are shown in Table 2.
As anticipated, the PCB groupings were highly correlated with each
other and with the more highly chlorinated congeners.
Of the 15 congeners analyzed, four (PCB-28, PCB-52, PCB-105,
and PCB-128) were excluded from data analysis as fewer than 35%
of the study samples had values above the limit of detection
(Supplementary Table S1). The results of the adjusted analyses of
the other PCBs are displayed in Table 3. There were significant
inverse associations between TGCT risk and 8 of the 11 PCBs
analyzed: PCB-118 (Q4 OR, 0.55; 95% CI, 0.400.76; P trend = 0.0007),
PCB-138 (Q4 OR, 0.46; 95% CI, 0.320.66; P trend = 0.0001), PCB-153
(Q4 OR, 0.45; 95% CI, 0.310.66; P trend = 0.0003), PCB-156 (Q4 OR,
0.57; 95% CI, 0.400.81; P trend = 0.002), PCB-163 (Q4 OR, 0.59;
95% CI, 0.420.83; P trend = 0.001), PCB-170 (Q4 OR, 0.56; 95% CI,
0.390.80; P trend = 0.002), PCB-180 (Q4 OR, 0.56; 95% CI, 0.380.82;
P trend = 0.003), and PCB-187 (Q4 OR, 0.004; 95% CI, 0.420.86;
P trend = 0.004). When the analysis was restricted to the nonseminoma cases, the same eight congeners were significantly
inversely associated with risk. Among the seminoma cases, five of
the congeners were statistically significantly inversely associated
with risk, although the gradient of the trends tended to be less

steep than the gradient of the nonseminoma relationships.
The analyses of PCBs by functional groups are shown in Table 4.
Total TGCT was significantly inversely associated with all PCB
groupings examined: Wolff group 1B (P trend = 0.02); Wolff group 2A
(P trend < 0.0001); Wolff group 2B (P trend = 0.0002); Wolff group 3
(P trend = 0.002); the inducers of UDP-GT, CYP1A, and CYP2B
(P trend = 0.0005); the phenobarbital inducers (P trend = 0.0003); the
mixed function oxidase inducers (P trend < 0.0001); and the sum of
PCBs (P trend = 0.0004). Similarly, nonseminoma was significantly
inversely associated with all groupings, whereas seminoma was
associated with all groups except Wolff group 1B (P trend = 0.33).
As DDE had previously been shown to be related to increased
risk of TGCT in the STEED population, an examination of sum of
PCBs, stratified by DDE, was undertaken.
As shown in Table 5, none of the trends in total PCB exposure
was significant in the strata of individuals with low DDE levels.
Among the individuals with high DDE levels, however, PCB levels
were significantly inversely related to risk of all TGCT (P trend = 0.03)
and to nonseminoma (P trend = 0.04). The interaction analyses, however, did not find, however, that the relationship between PCBs and
TGCT differed significantly by DDE stratum.
To determine whether the congener-specific or the PCB
grouping analyses were affected by the inclusion of persons whose
interval between serum collection and diagnosis was <1 year,
subgroup analyses that eliminated the data from those individuals
were performed. The subgroup analyses resulted in no differences
in results (data not shown).
Discussion
The incidence of TGCT has been increasing in the United States
since before World War II (19). Although few risk factors have been
identified, several studies have reported that there is a pronounced
birth cohort effect on risk, suggesting that changes in exogenous
exposures may be related to the trend (20). One exogenous
exposure, endocrine-disrupting chemicals, including PCBs, has
been the subject of much speculation, as animal data suggested
1905
Cancer Res 2009; 69: (5). March 1, 2009
Cancer Research
Table 4. Adjusted relative risk of TGCTs by quartile of PCB groupings in the STEED study
Controls
n
All TGCT
n
Seminoma
OR* (95% CI)
Wolff group 1B (101, 187)

(potentially estrogenic, weak phenobarbital inducers)
Q1
227
214
1.00
64
Q2
229
193
0.88 (0.661.18)
77
Q3
228
156
0.64 (0.460.89)
76
Q4
228
171
0.65 (0.460.93)
95
0.02
P trend
Wolff group 2A (118, 156)
(potentially antiestrogenic, immunotoxic, dioxin-like, non-ortho and mono-ortho substituted)
Q1
229
244
1.00
75
Q2
227
156
0.59 (0.440.79)
61
Q3
228
189
0.62 (0.450.85)
102
Q4
229
147
0.42 (0.290.59)
75
<0.0001
P trend
Wolff group 2B (138, 170)
(potentially antiestrogenic, immunotoxic, limited dioxin activity, di-ortho substituted)
Q1
228
231
1.00
71
Q2
228
164
0.66 (0.490.90)
60
Q3
228
176
0.58 (0.420.81)
92
Q4
229
165
0.46 (0.310.67)
90
0.0002
P trend
Wolff group 3 (99, 153, 180, 183)
(phenobarbital, CYP1A, and CYP2B inducers)
Q1
228
233
1.00
75
Q2
228
161
0.63 (0.470.85)
60
Q3
228
171
0.57 (0.410.80)
86
Q4
228
170
0.49 (0.340.70)
892
0.002
P trend
UDP-GT, CYP1A, and CYP2B inducers (99, 101, 118, 153, 156, 180, 183, 187)
Q1
228
236
1.00
75
Q2
228
166
0.64 (0.480.87)
62
Q3
228
166
0.55 (0.390.76)
85
Q4
228
166
0.47 (0.330.68)
90
0.0005
P trend
Phenobarbitol inducers (99, 101, 153, 163, 180, 183)
Q1
228
234
1.00
74
Q2
228
166
0.65 (0.480.87)
68
Q3
228
172
0.58 (0.420.81)
82
Q4
228
162
0.46 (0.320.66)
88
0.0003
P trend
Mixed function oxidase inducers (118, 138, 156, 170)
Q1
222
233
1.00
73
Q2
234
173
0.63 (0.460.84)
66
Q3
228
175
0.54 (0.390.75)
90
Q4
229
155
0.41 (0.380.60)
84
<0.0001
P trend
Total PCBs (99, 101, 118, 138, 153, 156, 163, 170, 180, 183, 187)
Q1
228
235
1.00
75
Q2
228
163
0.62 (0.460.83)
58
Q3
228
169
0.55 (0.400.77)
92
Q4
228
167
0.46 (0.320.67)
87
0.0004
P trend
Nonseminoma
OR* (95% CI)
OR* (95% CI)
1.00
0.97 (0.641.48)
0.77 (0.491.22)
0.80 (0.491.29)
0.33
150
116
79
76
1.00
0.83 (0.591.16)
0.56 (0.380.83)
0.55 (0.360.84)
0.004
1.00
0.64 (0.420.99)
0.83 (0.541.28)
0.44 (0.270.72)
0.002
169
95
87
71
1.00
0.56 (0.400.79)
0.50 (0.340.72)
0.38 (0.250.58)
<0.0001
1.00
0.70 (0.451.08)
0.75 (0.481.17)
0.49 (0.290.83)
0.01
160
104
84
74
1.00
0.68 (0.480.95)
0.49 (0.330.73)
0.43 (0.270.68)
0.0004
1.00
0.63 (0.410.97)
0.64 (0.401.00)
0.49 (0.290.81)
0.02
158
101
84
78
1.00
0.64 (0.460.91)
0.53 (0.360.78)
0.46 (0.290.71)
0.003
1.00
0.67 (0.441.02)
0.64 (0.411.00)
0.50 (0.300.83)
0.003
161
104
80
76
1.00
0.65 (0.460.91)
0.47 (0.320.70)
0.44 (0.280.68)
0.03
1.00
0.74 (0.491.12)
0.62 (0.390.97)
0.47 (0.290.79)
0.006
160
98
89
74
1.00
0.61 (0.430.86)
0.55 (0.380.81)
0.43 (0.280.67)
0.0009
1.00
0.64 (0.420.99)
0.66 (0.421.03)
0.42 (0.250.71)
0.003
160
107
85
70
1.00
0.65 (0.460.91)
0.47 (0.320.69)
0.39 (0.250.61)
<0.0001
1.00
0.58 (0.380.90)
0.67 (0.421.05)
0.45 (0.270.76)
0.01
160
105
77
79
1.00
0.65 (0.460.92)
0.46 (0.310.69)
0.45 (0.290.71)
0.001
(Continued on the following page)
Cancer Res 2009; 69: (5). March 1, 2009
1906
Table 4. Adjusted relative risk of TGCTs by quartile of PCB groupings in the STEED study (Contd)
Controls
n
All TGCT
Seminoma
Nonseminoma
OR* (95% CI)
OR* (95% CI)
OR* (95% CI)
224
171
175
162
1.00
0.88 (0.671.16)
0.73 (0.540.98)
0.61 (0.430.86)
0.006
74
60
91
88
1.00
0.90 (0.601.35)
0.89 (0.591.34)
0.64 (0.411.02)
0.05
150
111
84
73
1.00
0.84 (0.611.15)
0.62 (0.430.88)
0.55 (0.370.83)
0.007
Total PCB exposure

Q1
228
Q2
227
Q3
227
Q4
228
P trend
*Adjusted for matching variables, serum DDE level, age at serum draw, BMI, and height.
cTotal PCB exposure is sum of all PCB observations above the limit of detection for all PCBs assayed: 28, 52, 99, 101, 118, 128, 138, 153, 156, 163, 170,
180, 183, and 187.
that they might be related to a variety of male reproductive

disorders in humans (8, 20). The results of the current study,
however, that PCBs are inversely related to risk of TGCT do not
support the hypothesis. In that the inverse association seemed
more pronounced, although nonsignificantly so, among persons
with high serum levels of p,p-DDE, it is possible that the effect of
PCBs will depend on the mix of other endocrine disruptors also
present. Even at lower serum p,p-DDE levels, however, there was
no indication that PCBs served to increase risk.
PCBs are a group of related compounds composed of two carbonlinked benzene rings to which are attached between 1 and 10
chlorine atoms. Between 1929 and 1977, PCBs were manufactured
and used in the United States as insulators and coolants in electrical equipment, and in the production of numerous household
products. Concerns about possible long-term health effects first
surfaced in the 1960s when PCBs were reported to be prevalent in
wildlife and persistent in the environment (21). Subsequent animal
studies reported that PCB exposure resulted in several outcomes,
including neurobehavioral changes, hypothyroidism, reproductive
disorders, and tumors. Among humans, mass PCB food poisonings
in Japan (1968) and Taiwan (1979) resulted in chloracne, menstrual
irregularities, altered immune responsiveness, and general fatigue.
In 1977, the sole manufacturer of PCBs in the United States, the
Monsanto Company, ceased production 2 years before the formal
ban by U.S. Environmental Protection Agency. Based on the animal
data and on occupational studies in humans, IARC has classified
PCBs as being a probable carcinogen in humans (22).
In humans, PCBs are stored in adipose tissue and levels tend to
increase with age (12). Thus, it is difficult to determine in the
present study when the study participants were exposed. PCBs,
however, can cross the placental barrier and are present in breast
milk; thus, some of the exposure may have occurred in utero and/or
via breast-feeding. If significant exposure occurred in utero, PCBs
may also affect the risk of the male reproductive congenital
anomalies that are associated with TGCT. To date, very few studies
of either cryptorchism or hypospadias have examined PCB levels,
however. Two studies of cryptorchism and PCBs have been reported
and neither has found a relationship (23, 24). In contrast, a study of
the prevalence of hypospadias in Greenland found an unexpectedly
low rate despite high levels of PCBs in the population (25),
suggesting that PCBs might be inversely associated with risk
of hypospadias.
Among the testicular dysgenesis syndrome disorders (9) that

become evident in adulthood (impaired spermatogenesis and
TGCT), more studies have examined the relationship of PCBs with
the former than the latter. In general, the results of the PCB-fertility
studies are somewhat equivocal. Several studies have found
statistically significant associations with impaired sperm variables
(2628), whereas others have found associations only in subsets of
their populations (2931) or not all (32). In contrast, several studies
have reported direct associations between PCB levels and fertility
(33, 34). The summary of the international INUENDO study of PCBs
and fertility in four populations, however, concluded that a
representative congener, PCB-153, did not seem to affect fertility
or to have direct hormone-like activity (35).
Only one prior study of TGCT and PCBs has been reported to
date (13). A case-control study of Swedish men found no
association of TGCT with sum of PCBs, estrogenic PCBs, or
enzyme-inducing PCBs. An examination of PCB levels in the
mothers of the men, however, found that mothers of the cases had
significantly higher levels of 21 of the 37 congeners tested. When
the PCBs were examined by functional group, the analysis found
that the case mothers had significantly higher levels of sum of PCBs
and enzyme-inducing PCBs. Interpretation of the mothers results
is rather difficult, however, as the mothers blood samples were
obtained f30 years after their sons were born. Body burdens of
PCBs in women are affected by weight changes, child bearing, and
lactation over time, so it is unclear to what extent the mothers
PCB levels were representative of their levels during pregnancy
with their sons. It is uncertain why the sons results differed from
the results of the current study, but the discrepancy may be related
to the small size of the Swedish study (n = 58 cases, 61 controls),
differences in the PCB mixtures used in each country, and timing
of the collection of blood samples, as population PCB levels have
declined since they were banned. The blood samples in the
Swedish study were drawn at a later time (19972000) than the
samples in the current study, suggesting that the PCB levels might
be lower in the Swedish study. A comparison of median levels
of estrogenic PCBs and enzyme-inducing PCBs, however, only
partially supports this conjecture. The median level of estrogenic
PCBs among the Swedish control men was 25 ng/g lipid in contrast
with the median control level in the current study of 69.7 ng/g lipid.
The median level of the enzyme-inducing PCBs, however, was
174 ng/g lipid among Swedish controls and 73.9 ng/g lipid among
1907
Cancer Res 2009; 69: (5). March 1, 2009
Cancer Research
Table 5. Risk of TGCT by DDE and PCB levels in the STEED study
Low DDE (<147 ng/g lipid)

Cases/controls (n)
Sum of PCBs Q1
Sum of PCBs Q2
Sum of PCBs Q3
Sum of PCBs Q4
P trend
High DDE (z147 Ag/g lipid)
Cases/controls (n)
Sum of PCBs Q1
Sum of PCBs Q2
Sum of PCBs Q3
Sum of PCBs Q4
P trend
P interaction
All TGCT
Seminoma
Nonseminoma
OR* (95% CI)
OR* (95% CI)
OR* (95% CI)
343/455
1.00
0.59 (0.410.85)
0.67 (0.421.06)
0.75 (0.401.41)
0.36
123/455
1.00
0.70 (0.411.19)
0.72 (0.371.41)
1.05 (0.452.44)
0.93
219/455
1.00
0.56 (0.370.85)
0.62 (0.361.07)
0.59 (0.271.28)
0.15
390/457
1.00
0.41 (0.200.85)
0.27 (0.130.56)
0.28 (0.130.60)
0.03
0.48
189/457
1.00
0.33 (0.130.87)
0.35 (0.140.91)
0.27 (0.100.74)
0.11
0.29
201/457
1.00
0.45 (0.201.00)
0.21 (0.090.48)
0.28 (0.120.66)
0.04
0.72
*Adjusted for reference age, race, serum date, family history of TGCT, cryptorchism, age at serum draw, BMI, and height.
U.S. controls. It is also possible that the level of risk or protection of

PCBs is determined by the mixtures of congeners present in any
location or the presence of other compounds with endocrinedisrupting properties. For example, p,p-DDE was not related
to TGCT in the Swedish study but was related to risk in the
STEED population (10). This difference may be explained by
the lower general level of p,p-DDE in Sweden than in the United
States (36).
Why PCBs would be inversely associated with TGCT remains to
be determined. p,p-DDE, which has shown antiandrogenic
properties, was associated with increased risk in the same
population. PCBs have a range of estrogenic, antiestrogenic,
androgenic, and antiandrogenic effects (12). Examining the effect
by the Wolff groupings (15), only two (PCB-101 and PCB-187) of the
PCBs in Wolff group 1 (potentially estrogenic) were examined in
the current study. Whereas PCB-101 had no relationship with risk,
PCB-187 was inversely associated with both TGCT and nonseminoma. All of the Wolff group 2 (potentially antiestrogenic,
dioxin like) PCBs examined (PCB-118, PCB-156, PCB-138, and PCB170) were significantly inversely related to TGCT risk. Two of the
four Wolff group 3 (phenobarbitol, CYP1A, and CYP2B inducers)
PCBs were significantly inversely related to risk (PCB-153 and
PCB-180), whereas there was no relationship with PCB-99 or PCB183. Given that there were inverse relations in all the groups, it is
not clear what effect is most important in determining risk. In light
of the DDE relationship, however, it seems unlikely that an
antiandrogenic effect of PCBs would be inversely related to risk. An
antiestrogenic effect, however, might be related, especially as the
ratio of androgenic to estrogenic exposures has been postulated to
be important in TGCT development. Although there was no
statistically significant difference in the relationship of PCB levels
to TGCT in low and high DDE strata, significant trends only in the
high DDE strata may indicate that the presence of other endocrine
disruptors is critical to determining the effect of PCBs.
A major advantage of the current study was that prediagnostic
serum samples were analyzed. Other advantages were that
Cancer Res 2009; 69: (5). March 1, 2009
participants were drawn from a well-defined population, the

tumors were histologically confirmed, and the participants were
likely to be representative of a wide spectrum of the underlying
population. Study limitations include that some potential participants could not be contacted due to deployment, the analysis
adjusted for self-reported body size rather than measured body
size, and the study could not determine when and how the
participants were exposed to PCBs. Inability to contact men due to
deployment presents a potential bias in that deployed men might
be different in some way that nondeployed men. As most young
men in military service are healthy and fit, however, it would seem
unlikely that that deployment would confer substantial bias.
In conclusion, the current study suggests that PCBs are inversely
associated with the risk of TGCT, particularly with nonseminoma.
The results argue for further examination of PCBs and TGCT in
other populations, as PCBs are detectable in a large proportion of
the worlds population. It will be particularly instructive to examine
PCB levels in concert with other endocrine disruptor levels to
determine whether the inverse association detected in the current
study is a result of exposure to other environmental chemicals.
Disclosure of Potential Conflicts of Interest

No potential conflicts of interest were disclosed.
Acknowledgments
Received 10/9/2008; revised 12/5/2008; accepted 12/8/2008; published OnlineFirst
02/17/2009.
Grant support: Intramural Research Program of the National Cancer Institute,
NIH, Department of Health and Human Services.
The costs of publication of this article were defrayed in part by the payment of page
charges. This article must therefore be hereby marked advertisement in accordance
with 18 U.S.C. Section 1734 solely to indicate this fact.
We thank Emily Steplowski and Leslie Carroll of IMS for their contributions in data
management and analysis and the STEED participants, without whom there would
have been no study.
The opinions or assertions contained herein are the private views of the authors,
and are not to be construed as official, or as reflecting the views of the U.S.
Department of the Army or the U.S. Department of Defense.
1908
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NIH Public Access

Author Manuscript
Gefahrst Reinhalt Luft. Author manuscript; available in PMC 2011 June 13.
Published in final edited form as:

Gefahrst Reinhalt Luft. 2011 January ; 71(1-2): 2532.
Polychlorinated Biphenyl (PCB) carcinogenicity with special

emphasis on airborne PCBs
Larry W. Robertson1,2 and Gabriele Ludewig1,2
1 Interdisciplinary Graduate Program in Human Toxicology, University of Iowa
2
Department of Occupational & Environmental Health, University of Iowa
Abstract
Polychlorinated biphenyls (PCBs) are industrial chemicals used in various applications requiring
chemical stabilityand have now become widely dispersed. Their characteristics of persistence, low
water/higher lipid solubility, contribute to their ability to bioconcentrate and bioaccumulate.
Traditionally PCBs have been regulated as food contaminants and the general population is
primarily exposed by that route. PCBs in foodstuffs are generally higher chlorinated, resistant to
metabolic breakdown, and elicit toxic changes that are thought to be predominantly receptor/
parent PCB-driven. But for certain occupational exposures, and for those persons residing or
working in contaminated buildings, and in large cities, an inhalation route of exposure may
predominate. Airborne PCBs are, in contrast to foodborne PCBs, lower chlorinated, more volatile,
and subject to metabolic attack. In this review, we have explored (geno-) toxic manifestations of
PCBs typical of those found in air. Here metabolic conversion of the parent PCB to hydroxylated
and other metabolic progeny appear to play a dominant role, especially in genotoxicity. We should
be cognizant of the impact of exposures to airborne PCBs for those individuals who are
occupationally exposed, for persons living near contaminated sites, for those who work or go to
school in contaminated buildings, and especially cognizant of the young, the socio-economically
disadvantaged and medically-underserved or nutritionally-deficient populations.
1. Introduction - Sources of PCBs
PCBs have been commercially manufactured since the 1920s for use as dielectrics in
transformers and capacitors, as cooling fluids in hydraulic systems, in the formulation of
lubricating and cutting oils, in pesticides and flame retardants, and as plasticizers in paints,
copying paper, adhesives, sealants and plastics [1,2]. The stability of these compounds, one
of their commercial attributes, has led to their worldwide distribution in the environment, as
first reported by Jensen in 1966 [3]. The production of PCBs peaked in the 1970s and has
steadily declined thereafter as many countries throughout the world have banned their use or
limited their production. Nevertheless these compounds remain in use today in our
environment and represent a potential human health hazard [46].
An overlooked source of PCB exposure is airborne PCBs. Regular monitoring of
environmental PCBs in water, fish, and sediment of the Great Lakes and other regions in the
US started in the 1980s [7]. Such non-atmospheric sources of PCBs are carefully monitored
Larry W. Robertson, Ph.D., M.P.H., Director, IDGP in Human Toxicology, Professor, Department of Occupational and
Environmental Health The University of Iowa, College of Public Health, 100 Oakdale Campus #219 IREH, Iowa City, Iowa
52242-5000, Phone: 319-335-4554, Fax: 319-335-4290, larry-robertson@uiowa.edu
Gabriele Ludewig, Dr. rer. nat., Associate Professor, Department of Occupational and Environmental Health The University of Iowa,
College of Public Health, 100 Oakdale Campus #214 IREH, Iowa City, Iowa 52242-5000, Phone: 319-335-4650, Fax: 319-335-4290,
gabriele-ludewig@uiowa.edu
Robertson and Ludewig
Page 2

and regulated. Air as a source of PCB exposure, however, was nearly completely ignored
until a decade ago. Systematic measurements of atmospheric PCBs started only in the
1990s. The first urban monitoring site in the USA was installed in Chicago in 1995. The
level of PCB contamination in the air is strongly influenced by temperature. In Chicago air
concentrations between 100300 pg/m3 in winter and up to 5,00016,000 pg/m3 on hot
summer days were reported (reviewed in [8]). The levels at the sources of these atmospheric
PCBs may even be significantly higher. Inhalation exposure is considered to be a major
route of occupational exposure to PCBs, and it was estimated that in capacitor workers, for
example, a maximum of 80% of adipose PCBs may have been absorbed by inhalation
exposure [9]. Recently high levels of PCBs were measured in indoor air in buildings
constructed in the 1970s using joint sealants that contained 49% PCBs. Indoor air
concentrations up to 13,000 ng/m3 were measured in some classrooms of a contaminated
school [10], which is more than an order of magnitude above the NIOSH guidelines of 1 pg/
m3 for occupational settings. Other possible sources for indoor PCBs are believed to be data
screen terminals [11], ceiling tiles and fluorescent lights [12]. It was reported that the
concentration of PCBs in indoor air can be at least an order of magnitude higher than
outdoor air [1315], however, regional outdoor levels can be very high due to activities like
building renovations, dredging, or contamination from cement factory exhaust [1618]. The
serum levels of workers engaged in sealant removal was 210 times higher at the end of
these activities than they had been one year before [19]. Thus under certain circumstances
the intake from inhalation exposure exceeds PCB intake from food.
PCBs in foods, like fish or mothers milk, and in human adipose tissue are usually the higher
chlorinated ones, where congeners like PCBs 153, 180, 183 and others predominate.
Airborne PCBs are very different, since they require volatilization. Major congeners in
Chicago air, for example, are lower congeners, like PCBs 4, 8, 11, 18, 28, 52, 95, 112, to
name some (Figure 1) [20]. As a consequence of this difference, in two populations in Italy
the more urban group had significantly higher levels of lower chlorinated PCBs (PCB52 was
about 100-fold higher) than the population in a more rural environment [21]. In Germany,
PCB28 and PCB52 were the prevailing congeners in indoor air of contaminated schools
[10,22]. Elevated levels of PCB28 and PCB52 were measured in the blood of teachers from
these schools compared to non-contaminated schools, whereas the mean blood levels of
higher chlorinated PCBs, i.e. PCB138, 153 and 180 were almost identical [22]. Children in
schools with 690 20,800 ng PCB/m3 air had median levels of 6, 9, and 5 ng/l PCB28, 52,
and 101, respectively, whereas children in non-contaminated schools had levels below the
detection level of 1 ng/L [23]. Both groups had no significant differences in PCB138, 153
and 180 levels, indicating that indoor air exposure contributed to the PCB body burden. In
Germany the non-occupational tolerable indoor air PCB concentration was set at 300 ng/m3
based on a tolerable daily intake (TDI) of a total of 1 ug/kg body weight [24]. Not only were
these levels exceeded in several schools, but this TDI is also based on a chronic toxicity
study with a commercial PCB mixture, which measured hepatic enzyme induction as
endpoint [25]. Airborne PCB profiles are distinctly different from those of commercial PCB
mixtures, like Aroclor 1254, and enzyme induction in the liver is an inappropriate endpoint
of toxicity for inhalation exposure of airborne PCBs.
2. Carcinogenicity of PCBs
PCBs have been categorized by the International Agency for Research on Cancer (IARC) as
Probably carcinogenic to humans (Group 2A) [26], and by the National Toxicology
Program 11th Report on Carcinogens as Reasonably anticipated to be human carcinogens.
A detailed summary of the health effects of PCBs was prepared for the Agency for Toxic
Substances and Disease Registry (ATSDR) and published in 1990 as a Toxicological Profile
[27]. Other detailed reviews are available in the literature, emphasizing animal data, human
Page 3
studies, and mechanistic studies of the genotoxic effects of PCB individual congeners and
mixtures [1,28,29]. A summary of recommendations for selected IARC-classified agents
was recently published [30,31].
3. PCBs are Initiators and Promoters of Carcinogenesis
The process of carcinogenesis in many, if not all, tissues involves at least two stages,
initiation and promotion, based on observations originally seen in mouse skin and later
demonstrated in other tissues [32,33]. Mechanistically, an initiator has been defined as an
agent which irreversibly alters the DNA sequence (i.e. a genotoxic compound), whereas a
promoter has been defined as an agent that alters the expression of genetic information
(epigenetic changes) in the cell [33]. Although their potency varies, the various commercial
halogenated biphenyl mixtures have been uniformly reported to have promoting activity in
various liver models [29,34]. It had been widely assumed that PCBs do not have initiating
activity, but in several studies Oesterle and Deml consistently found a small number of
enzyme altered foci in livers of rats treated only with the PCB mixture Clophen A50
(reviewed in [35]). The same was noted for the brominated biphenyl mixture fireMaster
BP-6 [36]. Hayes and coworkers had used the modified Solt-Farber initiation-selection
protocol to investigate the initiating activity of several PCBs and mixtures [37], with
negative results. Our mechanistic studies predicted that the choice of PCBs in the Hayes
study was not fortuitous, because the selected PCBs were so slowly metabolized that
detection of initiating events was precluded. Our data, however, clearly show that several
lower chlorinated PCB congeners and two PCB3 metabolites are positive in this initiation
assay [38,39]. In addition, the brominated congener 3,3,4,4-tetrabromo-biphenyl (PBB77)
was found to have both, initiating and promoting activity [40]. Because the long term
administration of PCB mixtures and certain congeners leads to the development of hepatic
tumors, PCBs by definition have both initiating and promoting activities. Besides liver
tumors, several PCB mixtures and congeners were reported to also promote lung tumors in a
mice model [41,42] and to cause lung tumors in long term rat studies conducted by the
National Toxicology Program (NTP), something to keep in mind in connection with
airborne PCBs. PCBs are classified as proven animal carcinogens and probable human
carcinogens. The risk of airborne exposure for human health is just beginning to receive the
attention that it deserves. It is the intent of the authors to devote the space below to issues
related to lower chlorinated PCBs, as predominate in air, and their adverse effects,
especially related to genotoxicity and carcinogenicity.
4. Metabolic Activation of Lower Chlorinated PCBs to Reactive

Intermediates
It has long been recognized that biphenyl and halogenated biphenyls, particularly the lower
chlorinated congeners, are hydroxylated in vivo and in vitro (see review by [43]). These
hydroxylation reactions are primarily catalyzed by isoforms of cytochrome P-450. Our
experiments with PCB3 (4-chlorobiphenyl) and rat liver microsomes showed that five
mono- and three di-hydroxy metabolites were formed [44]. The metabolism of PCB3 by
cytochrome P-450 probably involves an arene oxide intermediate [43,45]. Other arene
oxides could be involved in the oxidation of the mono- to the di-hydroxy forms. Arene
oxides are strong electrophiles which may react with critical cellular targets. The
dihydroxybiphenyls may be further oxidized by various enzymes like peroxidases [46],
prostaglandin synthase [47] and cytochrome P-450s to the corresponding quinones with the
formation of a semiquinone intermediate. Ortho- and para-quinones are formed from diOHPCB3 in vitro and demonstrate high reactivity toward nitrogen and especially sulfur
nucleophiles [48]. Other experiments demonstrated that the microsomal metabolism of
PCB3 resulted in the formation of adducts with nucleotides in vitro, preferentially with
Page 4
purines rather than pyrimidines [45]. Most likely at least 1 of the 4 adducts seen is derived
from an arene oxide intermediate, the 3 other adducts after further oxidation probably from a
quinone [45]. These results suggest that several metabolic pathways and chemical species
could be involved in PCB-induced DNA adduction, without, however, telling us exactly
which enzymes or metabolites are involved.
5. PCBs as Mutagens and Genotoxins
The formation of PCB-DNA adducts was also reported in cellular systems [49], and studies
in vivo demonstrated covalent binding of radiolabeled PCB metabolites to nucleic acids
[5052]. The assumption that PCBs are not mutagenic is based on very little (and often
contradictory) data. Of 209 possible congeneric PCBs, only five congeners, one bromo-, a
few fluoro-biphenyls and five commercial PCB mixtures have been tested in the Ames
bacterial mutagenicity test. Most tests gave negative results, however, Aroclor 1221, 4bromobiphenyl, and some lower fluorinated biphenyls were positive [53]. The mutational
spectra gave evidence for two mutagenic species. The reported positive effects with PCB3
[54] were not confirmed by Schoeny [55]. Thus lower halogenated biphenyls may or may
not be mutagenic in bacteria, but it should be considered that the Ames test is insensitive
towards whole groups of compounds, like DES, benzene and reactive oxygen species
(ROS). One recent publication reported a small, but significant increase in lac I mutations in
livers of transgenic (BigBlue) mice after exposure to Aroclor 1253 [56]. Clearly this and
other mutagenicity tests, including those that measure chromosome mutations and genome
changes, and an analysis of more congeners is needed.
Recently we have shown that several PCB3 metabolites induce gene mutations,
chromosome breaks, chromosome loss and polyploidization in cells in culture and we
provided the first evidence that a PCB congener is mutagenic in vivo. As seen, the early
attempts to measure the genotoxicity of PCBs did not account for bioactivation of lower
chlorinated congeners. Clearly, under the right metabolic conditions certain lower
chlorinated PCBs can be bioactivated to genotoxins. The questions remain, which are the
active metabolites, which kinds of genotoxic damage are important, what are the
mechanisms of these genotoxicities, and are these issues really relevant for inhalation
exposure situations as those in our inner cities and contaminated buildings?
A series of PCB3 metabolites were tested in various genotoxicity assays to determine their
activity and genotoxicity profile. Both, the 3,4- and particularly the 2,5-quinone were very
efficacious and potent inducers of gene mutations at the HPRT locus (Table 1). Neither of
the corresponding dihydroxy metabolites nor the phenols had any activity in this assay. The
2,5-quinone (2,5-pQ) was also by far the most potent and efficacious inducer of
chromosome breaks as determined by Crest-negative micronuclei induction. This suggests
that at least some of the HPRT gene mutations may be due to breaks in the X-chromosome.
The ortho-quinone (3,4-oQ), 3,4- (3,4-Cat) and 2,5-dihydroxy (2,5-HQ) and 4-monohydroxy
metabolites (4-OH) induced some chromosome breaks at the highest concentration tested,
but their by far stronger activity was the induction of chromosome loss (Crest-positive
micronuclei). In this respect 4-OH and 2-OH were the most potent metabolites tested, while
the dihydroxy and quinone metabolites produced significant chromosome loss at more than
10-fold lower concentrations (above results were published in [57]. Two unique effects
stand out: only one metabolite, the 3,4-catechol, induced sister chromatid exchanges (SCE),
and only the 2,5-hydroquinone (2,5-HQ) caused tetraploidization of cells, and this with an
efficacy of nearly 100% at 7.5 M concentration [58]. Tetraploidization followed by uneven
chromosome loss is believed to be a major pathway of carcinogenesis. The mystery
surrounding this polyploidization is, however, the mechanism by which it occurs. According
to the staining pattern (all dark) the cells were not in the second M-phase as they should be.
Page 5
Time-lapse microscopy showed that these cells were arrested in first G2/M, but then went
out of G2/M and shortly afterwards fused with surrounding cells. This intriguing observation
is under further analysis.
Very interesting also is the fact that 2,5-HQ and its oxidation product 2,5-pQ have such
different profiles. To gain further insight into the mechanism and potential organ specificity
of effects, two cell lines were employed that differ in their amount of myeloperoxidase
(MPO), an enzyme found in bone marrow cells and others and expected to oxidize
hydroquinones to quinones. The 2,5-pQ induced DNA strand breaks, measured with the
COMET assay, in both cell lines and at 37 and 6C; The 2,5-pQ also increased intracellular
ROS while decreasing GSH levels. On the other hand 2,5-HQ induced COMETs and ROS
only in MPO-positive HL-60 cells and at 37C, indicating that this metabolite needs to be
bioactivated by MPO [59].
These results show that metabolites of PCB3 are indeed genotoxic and that each metabolite
induces its own, specific type of DNA damage. What these results do not explain is the
mechanism of genotoxicity for the individual endpoints, whether this is of any importance in
vivo or which metabolic activation pathway(s) could be leading to these effects. To address
these questions, male transgenic BigBlue rats were injected ip with PCB3, 4OH-PCB3, 3methylcholanthrene (3-MC), or corn oil and the induction of point mutations was analyzed
in the lacI indicator gene. PCB3 increased the mutation frequency in the liver (significantly)
and lung (non-significantly) of BigBlue rats, and changed the mutation spectrum in both
organs from predominantly transitions to predominantly to GC TA transversions. 4OHPCB3 had a similar, but smaller and effect that was below the level of statistical significance
[60,61]. Female rats were by far less susceptible to mutation induction in the liver,
reminding us that gender differences have to be considered [62] This demonstrates that this
PCB congener is mutagenic in vivo in the target organ liver and most likely also in the lung.
However, this still does not explain the mechanism of genotoxicity (DNA adduction or
ROS?), the metabolic activation pathway (ortho- or para-quinone, or epoxide or other
metabolite?), or whether the most likely route of exposure, inhalation of contaminated
indoor or outdoor air, may pose a significant risk for carcinogenicity in humans.
6. Oxidative DNA Damage
There is considerable evidence that lower chlorinated PCBs produce reactive oxygen species
(ROS) and intracellular oxidative stress [46,63]. Free radicals, particularly hydroxyl
radicals, may produce 8-oxodeoxyguanosine (8-oxodG), a DNA lesion that is highly
mutagenic, producing G T transversions [64]. Hydroxyl radicals can also attack fatty
acids (linoleic acid, linolinic acid, oleic acid, etc) and form lipid peroxidation-derived enals,
such as acrolein, crotonaldehyde, trans-4-OH-2-nonenal (4-HNE), and malondialdehyde
(MDA) [65]. These products can then modify DNA bases, resulting in cyclic adducts by
interaction of their difunctional groups with NH2 group in dA, dG ordC residues in DNA
[6668]. These cyclic adducts are mutagenic, producing base substitutions and deletions, for
example G T mutations from propano-dG and C A mutations from various etheno
adducts [64,69,70]. Therefore the question of mutagenicity of PCBs, especially congeners
that are prone to metabolic activation to redox cycling intermediates, like those from
airborne PCBs, should be re-analyzed.
7. PCBs cause Karyotype Changes

Induction of chromosome aberrations and genome mutations may be an essential part of
PCB carcinogenicity. Aroclors or Kanechlors produced chromosome aberrations in embryos
of PCB-treated ring doves [71] and in bone marrow cells of mice in vivo [72], and
chromosome aberrations in lymphocytes [73] and sarcoma cells in vitro [72]. Sargent and
Page 6
coworkers [73] also tested two congeners, 3,3,4,4-tetrachlorobiphenyl (TCB; PCB77) and
2,2,5,5-TCB (PCB 52) and found chromosome breakage and rearrangements with both and
a more-than-additive effect in combinations of the two PCBs in human lymphocytes in vitro.
Both congeners also induced preneoplastic foci and chromosome aberrations in rats in vivo,
and the combination of the congeners again had a more than additive effect [74,75]. The
most common types of aberrations were trisomy of chromosome 1 on its long arm and
monosomy of chromosome 3 on its short arm. Specific chromosome aberrations and
hepatocellular carcinoma correlated so that they hypothesized that genes involved in the
development of hepatic carcinoma may reside in chromosome 1 and/or 3 of the rat [75].
These data indicate that we need to better understand the mechanism of these effects and
structure-activity relationships for all PCB congeners, if we want to understand the cancer
risks associated with daily inhalation exposure to PCB congeners that may be bioactivated to
clastogens and aneugens. Our experiments with PCB3 metabolites are a first step in that
direction, but more work needs to be done.
8. PCBs cause Telomere Shortening
Telomeres are small but very important segments of chromosomes that are needed for
chromosome stability. PCB3 and the two metabolites tested, 2,5-quinone and 3,4-diOH
catechol, cause telomere shortening [76]. The effect was strong in TERT-immortalized
human primary fibroblasts and immortal keratinocytes (HaCaT) and not significant in
human primary fibroblasts, possibly because senescence started to interfere at the end of the
experiment. Shortening of telomeres would indicate that cells exposed to these compounds
may senesce prematurely in animals/humans.
Also measured was telomerase activity in cells exposed for 6 h to PCB3-2,5pQ and
PCB3-3,4diOH. A significant increase in activity in HaCaT and TERT-immortalized
fibroblasts was found. PCB3 did not increase the telomerase activity in HaCaT at the very
high concentration tested. These findings seem to contradict each other, since an increased
telomerase activity should prevent telomere shortening, which was observed with the same
compounds after 6 and 12 weeks of exposure. However, telomerase activity was measured
early, after only 6 h of exposure, and may not be maintained for long and of course, why and
how was telomerase activity increased: a feed-back mechanism signaling that the telomerase
is not functioning correctly, a consequence of signaling pathway activation, a consequence
of transient c-myc or growth factor activation?
Another puzzling aspect is the fact that not only the quinone, but also the mother compound,
PCB3 itself, reduced telomere length. The quinone was expected to produce intracellular
oxidative stress, a factor known to produce telomere shortening [77]. In addition, the
quinone can bind to DNA (and presumably RNA) and to proteins [45,48], which could
interfere with the telomerase function. The 3,4diOH metabolite is expected to be oxidized
with the formation of ROS and reactive semiquinone-quinone. PCB3 on the other hand is
not very reactive. Since these first observations we now have evidence that several
additional PCB congeners cause a shortening of the telomeres and, during long-term
exposure (14 weeks), to a reduction in telomerase activity in immortal human HaCaT
keratinocytes (Senthilkumar et al, unpublished results). Since stem cells and the basal cells
of permanently proliferating tissues like skin and GI tract express and depend on telomerase
activity [7880], a chronic exposure to these PCB congeners could have so far unrecognized
consequences.
Overall these new observations are extremely interesting; they pose a number of important
questions that need to be answered: what is the mechanism of telomere shortening by these
different compounds?; what is the cause of telomerase activity increase or decrease and how
Page 7
persistent is it?; which PCB congeners and metabolites have this effect ? and can we deduce
a structure-activity relationship?, which in tern may help us to unravel the mechanism(s).
Obviously we are only at the beginning of extended series of experiments that are needed to
understand the risk that PCBs may cause through this newly discovered mechanism. If we
understand the effects of PCBs on the telomerase complex (hTERT and its RNA template,
hTR) we will be better equipped to understand the need to include telomere/telomerase
research in the analysis of the effects of many man-made compounds.
9. Exposure of Humans to Airborne PCBs
The importance of airborne PCBs is now becoming understood. A PubMed search with the 2
keywords PCB and air produced 136 hits for the three most recent years (20072009)
alone, more than in the three previous decades (1970s 90s) together. Little is known,
however, about the toxicity of these airborne PCBs and the consequences of exposure by
inhalation compared to ingestion. Airborne PCBs are lower chlorinated and therefore
relatively easily metabolized. This results in low levels of those PCB congeners detectable
in blood, but at the same time provides bioactivated intermediates. Although our daily
exposure to these airborne PCBs may be low under most circumstances, children playing
near Superfund sites in hot summer days, workers moving dried dredging material or
demolishing buildings containing PCBs, or families living unknowingly in buildings with
high indoor PCB concentrations, may be exposed to significant levels of these airborne
PCBs for extended periods of time. We should strive to understand the potential risks of
such exposure, and to understand the mechanisms of toxicity, so that we can devise
recommendations, protective strategies, predictions about possible susceptibility factors and/
or interactions with other compounds. Very often Superfund sites, contaminated buildings,
and multiple chemical exposures are found in poor neighborhoods, with medically
underserved and nutritionally deficient children. Understanding and, if needed, ameliorating
the risks is a matter of environmental justice and social responsibility.
Acknowledgments
Many of the studies referenced and cited in this short review were supported by funding from NIH (ES 013661, ES
05605). The opinions expressed are solely those of the authors, and do not reflect an official policy of the granting
agencies. The authors recognize that the research summarized here would have not been possible without the
dedicated hard work of graduate students, postdocs and staff. Their contributions are gratefully acknowledged.
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Page 12
Figure 1.
Average PCB concentration in Chicago air for the calendar year 2007. Data are depicted as a
weight percentage, and are taken from a study by Zhao et al, 2010 [20].


25
15
0.6
0.5
3,4-Cat
3,4-oQ
2,5-HQ
2,5-pQ
2.5
2.5
15
75
100
50
MN chromos. Loss1
7.5 (PP)
5 (SCE)
SCE or Polyploidy2
COMET @ 37C & 6C in HL-60 & Jurkat 0.1 (ROS ), 2.5 (GSH )
COMET @ 37C, not 6C, in HL-60, not in Jurkat 0.1 (ROS )
COMETS & Others (HL-60, Jurkat)3
Xie [59]
Flor [58]
Zettner [57]
MN: micronucleus assay; SCE: sister chromatid exchange assay; PP: polyploidy; TG: thioquanine. These data are associated with the following published studies:
75
4-OH-
2-OH-
PCB3
MN chromos. Breaks1
3-OH-
Gene mutations (TG-resist.)1
Compound
Genotoxic profile of PCB3 metabolites, 2-hydroxy-PCB3 (2-OH-), 3-hydroxy-PCB3 (3-OH-), 4-hydroxy-PCB3 (4-OH-), 3,4-dihydroxy PCB3 (3,4-Cat),
3,4-ortho quinone (3,4-oQ), 2,5-hydroquinone (2,5-HQ), and 2,5 para quionone (2,5-pQ). Numbers are LOEL (M) for that endpoint.
Table 1
Page 13
Polish Journal of Environmental Studies Vol. 13, No. 2 (2004), 203-208
Polychlorinated Biphenyls in the Sediments

of the Odra River and Its Tributaries
T
M. Rodziewicz, A. Kaczmarczyk, E. Niemirycz*
Institute of Meteorology and Water Management, Maritime Branch, Department of Water Pollution Control
ul. Jakowa Dolina 29, 80-286 Gdask, Poland
Received: 28 March, 2003

Accepted 18 July, 2003
Abstract
This paper presents the results of polychlorinated biphenyls (PCBs) determination in
sediments from different sites of the Odra River and its tributaries, collected in 1998-2002.
It was found that concentrations of PCBs varied significantly. The results of individual congeners show
that contents of PCBs 180 and 138 were the highest in the sediments investigated. For better identification
of places most polluted by PCBs, PCB contents on TOC (total organic carbon) basis in surface sediments
are presented.
Keywords: PCBs, TOC, sediments, the Odra River
Introduction
Polychlorinated biphenyls (PCBs) are persistent organic compounds of anthropogenic origin, which have
been produced on an industrial scale since 1929. They
were commonly used in the period 1950-1980. PCBs
have low or no flammability, good thermal stability,
low rates of evaporation and high electrical resistivity.
For all these reasons, they have been used in many industrial branches as heat transfer fluids, dielectric for
transformers and capacitors, additives in hydraulic fluids
in vacuum and turbine pumps, and plasticizers. They have
been incorporated into formulations for printing inks,
pesticides, paints, flame-retardants, waterproofing agents
and adhesives. PCBs enter the environment mainly during
accidents, not controlled or not properly controlled disposal of products containing the substances, chlorination
of drinking water and wastewater and as a by-product in
the chlorine bleaching of wood-pulp [1 - 8].
PCBs are stable compounds, resistant to decomposition by physical, chemical and biological factors. It
Corresponding author; e-mail: E.Niemirycz@imgw.gdynia.pl
is estimated that the half-life of PCBs in the environment is between 10 and 15 years [9, 10]. PCBs can
be removed from the natural environment by reaction
with active forms of metals and alkali, hydrogenation
as well as by photolysis, thermochemical reactions and
microbiological degradation [1]. The number and position of the chlorine atom in the molecule determine
PCBs rate of degradation. Mono and dichloro biphenyls are more easily decomposed than polychlorinated
biphenyls. The congeners containing the chlorine atoms
in one benzene ring are more easily degraded than the
ones with chlorine atoms in two rings. PCBs containing
a chlorine atom in orto position (in one or two benzene
rings) degrade with great difficulty [2].
PCBs are present in almost every element of the
environment because of their resistance to degradation, low rates of evaporation, lipophilicity and usage on a large scale for over 60 years (Table 1). High
concentrations of PCBs occur in cold and moderate
climates in the northern hemisphere. These substances
can be transported over large distances, and therefore
they are found in distant regions of Africa and Antarctica [1, 8, 11].
204
Rodziewicz M. et al.
Despite the relatively low concentrations of PCBs
in the natural environment, they have a negative influence on living organisms. PCBs undergo accumulation in consecutive links of the food chain. Plants,
crustaceans and fish easily absorb them. This results
in a decrease of photosynthesis, plant and fish growth
and reproductive capacity of aquatic organisms, and also
in increase of larvae mortality [13]. In the case of mammals, PCBs could cause pathologic changes in spleen and
skin. They also can damage immune, digestive, nervous
and reproductive systems, and promote the generation of
tumours [11, 14, 15, 16].
Within the International Odra Project (IOP) and
the individual project (No 6P04G02719) of the KBN
(Polish State Committee for Scientific Research), the
Department of Water Pollution Control, Maritime
Table 1. Polychlorinated biphenyls in the natural environment

[2, 9, 12].
Medium
Concentration
Air
0.1 20 ng/m3
Water
0.001 - 908 ng/l
Sediment
1.1 6000 g/kg (dry wt.)
Plankton
0.01 - 20 mg/kg
Invertebrates
0.01 - 10 mg/kg
Fish
0.01 - 25 mg/kg
Bird eggs
0.1 - 500 mg/kg
Human beeing
0.1 - 10 mg/kg
Table 2. Determined PCB congeners.

No
Number of PCBs acc. to IUPAC
TEF - Toxic Equivalency Factor
1.
52
2.
101
3.
118
0.0001
4.
153
5.
138
6.
180
7.
189
0.0001
Structural formula
205
Polichlorinated Biphenyls in the Sediments of Odra River...

Table 3. Parameters of chromatographic determination of PCBs.
System 1
System 2
Chromatograph type and

model
GC Hewlett Packard 6890
GC 8000 Fisons
Column
HP 5.30 m x 0.25 mm x 0.25 m
HP Ultra 2.25 m x 0.2 mm x 0.33 m
Detector
HP 5973 MSD in SIM mode
ECD
Injection temperature
300 oC
280 oC
Carrier gas
flow rate
60 C, step 15 C/min to 150 C, step 3 C/min to

300 oC
Helium
0.9 ml/min
Make-up gas
Nitrogen
Volume of injected sample
1l
1 l
Temperature program
Branch of the Institute of Meteorology and Water

Management carried out studies on especially toxic
organic substances in the river environment. This paper presents the results of PCB contents in sediments
collected from the Odra River and its tributaries in
1998-2002 [17, 18, 20].

Sediment samples were collected during five campaigns: in May and November 1998, June 1999, May/
June 2000 and June 2002. The location of sampling sites
in the Odra basin is presented in Figure 1 [17-18].
PCB congeners determined in collected sediment
samples are shown in Table 2.
80 oC, step 4 oC/min, to 280 oC

Helium
1 ml/min
Discussion of Results
The concentrations of 7 PCBs in sediment samples collected at different sites along the Odra River and its tributaries in May 1998 ranged from 1.3 g/kg (dry wt.) in the Odra
at Krosno Odrzaskie to 13.6 g/kg (dry wt.) in the Warta
near confluence. In November 1998, the sum of 7 PCBs
in all studied sediments varied from 1.3 to 28.0 g/kg (dry
wt.). The lowest concentration was measured in the Odra
for Widuchowa while the highest was in the Odra Braid.
Sample Preparation
Samples of surface sediments (0-5 cm) were collected
with a stainless steel bucket, transferred to glass vessels,
subsequently frozen, and stored at -20 C. After being defrosted, the samples were lyophilized, then sieved manually (sieve mesh 2 mm) and ground in a ball-grinder.
The internal standard (9-chloroanthracene) was added
to a 2 g sample of dry sediment and extracted with dichloromethane in Soxhlet apparatus for 15 h. The extract
was condensed by evaporation of the excess solvent in
evaporator under reduced pressure. After sulphur removal, achieved by adding copper powder, the extract was
purified on the micro-column filled with silica gel. A 9:
1 (v/v) mixture of hexane and dichloromethane was used
for elution. The eluate was condensed by evaporation of
the excess solvent in a stream of nitrogen and analyzed
chromatographically [17 - 18].
Chromatographic Analysis
The chromatographic analyses were performed on two
gas chromatographs. The chromatographic run conditions
in analysis of PCBs are shown in Table 3.
Fig. 1. Sediment sampling sites in the Odra River and its tributaries, 1998-2002 [17-18].
206
Fig. 3. Average concentrations of PCB congeners in sediments

from the Odra River and its tributaries, 1998 2002.
Fig. 2. Contents of PCBs in sediments from the Odra River and

its tributaries, 1998 2002.
Sediments from the Odra basin taken in 1999 and 2000

had higher contents of PCBs than in 1998. In June 1999,
the concentration of PCBs ranged from 12.6 (Szczecin) to
58.2 g/kg (dry wt.) (Police), and in May/June 2000 from
17.9 (Barycz) to 81.2 g/kg (dry wt.) (the Odra Braid).
In 2002 sediment samples were collected from the Odra
River at Police and Krajnik Dolny. At these sites the contents of 7 PCBs were 12.8 g/kg (dry wt.) and 14.2 g/kg
(dry wt.), respectively (Fig.2). Generally, by PCBs most
polluted the sediment was taken near the Odra mouth,
while the least polluted was from the Barycz where the
river joins the Odra.
The decree of the Minister of the Environment in September 2002 on quality standards for soil and ground determines maximum allowable concentrations of the sum
of 7 PCB congeners (No: 28, 52, 101, 118, 138, 153, 180)
in the surface layer of sediments accumulated or originating from the reservoirs of flowing or stagnant waters [19].
The permissible value of PCBs for protected, agricultural,
forest, urban and recreation-rest areas is 20 g/kg (dry
wt.) and for industrial and traffic areas 2000 g/kg (dry
wt.). The investigation of the PCB contents in sediments
indicates that permissible concentration levels of PCBs in
surface sediments of protected areas (non-polluted) has
been exceeded in 13% of samples from November 1998,
in 79% of samples from June 1999 and in 90% of samples
Fig. 4. Average concentrations of PCBs on TOC basis in sediments from the Odra river and its tributaries, 1998 2002.
from May/June 2000 [20-21]. In the case of sediments

from 1998 and 2002, PCB concentrations were below
permissible value.
PCBs 180 and 138 were found in the highest concentrations in the studied sediments (Fig. 3).
Data for sediments polluted with PCBs in the Odra
basin after the flood in the summer of 1997 are available
in literature. The concentration of 7 PCBs (No: 28, 52,
101, 118, 138, 153, 180) in sediment samples taken at different sites in the upper and middle Odra in August 1997
varied from below detection limit 0.05 g/kg (dry wt.) to
420 g/kg (dry wt.). The allowable concentration of PCBs
in surface sediments of protected areas (non-polluted),
207
Polichlorinated Biphenyls in the Sediments of Odra River...

i.e. 20 g/kg (dry wt.) was exceeded in 24% of examined
sediment samples [22].
As found by Konat and Kowalewska, the sum of
concentrations of 7 PCBs in sediments collected from the
Szczecin Lagoon before the 1997 flood was not higher than
20 g/kg (dry wt.); only at one station did 7 PCBs reach a
rather high value of ca. 55 g/kg (dry wt.) [23]. The highest
concentration of PCBs was observed not in the vicinity of
the most probable sources of pollution, but in areas of intensive sedimentation. One month after the flood of 1997, the
concentrations of 7 PCBs were about 1.5 - 3.7 times higher
than before this disaster. Even 2 years after the flood, PCB
pollution was still at a high level. These facts confirm the
conclusion drawn from our investigation, that an increase
of PCB concentration level occurred in sediment samples
taken from the Odra and its tributaries after the 1997 flood,
up to the year 2000 [23].
In many publications, for better identification of places
most polluted by PCBs, PCB concentrations on TOC basis in
surface sediments are presented (Fig. 4). The high concentration of PCBs on TOC basis in surface sediments (>2000 ng
PCBs/g TOC) were noted between the site where the Nysa
uycka joins Odra and Mescherin at the Odra River. These
values were much higher than presented in the earlier publication concerning the Szczecin Lagoon [24].
Summary
The studies on the PCB contents in sediments from

the Odra basin conducted in the 5-year period after a
disastrous flood in 1997 indicate that PCB compounds
occurred in variable concentrations. Most of the samples exceeded the permissible value of sum PCBs for
non-polluted areas (20 g/kg dry wt.) [19].
The highest contents of PCBs were found at Bytom Odrzaski, Mescherin, Szczecin and Roztoka
Odrzaska; this could indicate the discharge of PCBs
from industrial plants and rubbish.
Sediments from the Odra River in the region between
the Nysa uycka confluence and Mescherin can be
regarded as most polluted with PCBs (PCBs on TOC
basis). The PCB concentrations were significantly
higher than in sediments from the German zone of the
Szczecin Lagoon.
After the 1997 flood, up to the year 2000, an increase
of PCB concentrations in the studied sediments of the
Odra and its tributaries was observed. The similar
phenomenon was noted in the case of sediments from
the southern Baltic Sea.
Acknowledgements
This work was done within the framework of International Odra Project (No 423KfK9702) coordinated by
Professor Arndt Knchel from Hamburg University and
the individual project of the Polish State Committee for
Scientific Research (No 6P04G02719).
The authors are obliged to Elbieta Heybowicz and Regina Taylor of the Institute Meteorology and Water Management for their help in preparing this publication.
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fate, The Science of the Total Environment, 280, 1, 2001.
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25. KACZMARCZYK A., SOSNOWSKA M., NIEMIRYCZ
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193-201, 14-17 maja 2000, Wrocaw, 2001.

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Tugas Toksikologi dan Kesehatan Lingkungan

MEKANISME TERJADINYA KANKER OLEH PCBS

PROGRAM STUDI MAGISTER

1.2. Rumusan Masalah

Kanker dan Karsinogenesis

terpapar suatu karsinogen sampai terlihat kanker secara klinis.

Periode laten dari

pada hewan laboratorium/percobaan. Sebagai tambahan EPA mencurigai PCBs bersifat

Gambar 1. Struktur dasar kimia Polychlorinated Biphenyls

Sifat senyawa PCBs yang stabil dipandang sebagai kelebihan dalam

Gambar 2. Siklus penyebaran PCBs di lingkungan

Mekanisme Karsinogenisitas PCBs

Gambar 3. Proses mekanisme terjadinya kanker akibat PCB dan

Kerusakan Oksidatif DNA (Robertson & Ludewig 2011)

Gambar 4. Mekanisme Kerusakan Oksidatif DNA (Kira & Toren 2014)

NIH Public Access

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Published in final edited form as:

Development of a Synthetic PCB Mixture Resembling the Average

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Congener profiles of polychlorinated biphenyls (PCBs) found in environmental samples are

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Environ Int. Author manuscript; available in PMC 2011 November 1.

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2. Materials and methods

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Environ Int. Author manuscript; available in PMC 2011 November 1.

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Environ Int. Author manuscript; available in PMC 2011 November 1.

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3. Results and discussion

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Environ Int. Author manuscript; available in PMC 2011 November 1.

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Environ Int. Author manuscript; available in PMC 2011 November 1.

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Environ Int. Author manuscript; available in PMC 2011 November 1.

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Environ Int. Author manuscript; available in PMC 2011 November 1.

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Environ Int. Author manuscript; available in PMC 2011 November 1.

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Environ Int. Author manuscript; available in PMC 2011 November 1.

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Environ Int. Author manuscript; available in PMC 2011 November 1.

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