BIOLOGI MOLEKULER ONKOLOGI

Blok 19 - Onkologi (ONK)

dr. Henny E. S. Ompusunggu,M.Biomed

Dept. Biologi Sel dan Molekuler, Fakultas Kedokteran
Universitas HKBP Nommensen, Medan
 BIOLOGI MOLEKULER ONKOLOGI

Onkos (Yunani ) = tumor (massa)

Kanker merupakan kelainan genetik kompleks

yang melibatkan kelainan struktural dan
kelainan ekspresi gen

Penyebab: perubahan protein yang

mengkode protoonkogen dan tumor supresor
Multiple genetic changes underlie the development of cancer

Chromosomes 1 2 3 4
mutation mutations mutations mutations

Normal Malignant
cell cell

Four or more somatic mutations are usually required to

produce a cancer cell
 Klasifikasi tumor menurut sel embrionik asalnya :

1. Karsinoma
>80% tumor
Berasal dari jaringan endodermal/ektodermal
kulit, lapisan epitel organ internal, dan kelenjar
2. Kanker sel darah
Hematopoietic cells Leukemia dan Limfoma
3. Sarkoma
Jaringan ikat jaringan mesodermal tulang, lemak
dan tulang rawan
 SIKLUS SEL
Siklus sel merupakan fungsi sel yang paling
mendasar berupa duplikasi akurat sejumlah
besar DNA di dalam kromosom, dan kemudian
memisahkan hasil duplikasi tersebut hingga
terjadi dua sel baru yang identik.

Siklus sel yang berlangsung kontinu dan

berulang (siklik), disebut proliferasi
SIKLUS SEL
1. Fase G1 : 5-6 jam,
pertambahan ukuran sel,
sintesis RNA dan protein,
G1/S check point
2. Fase S : 10-12 jam, sintesis/
replikasi DNA
3. Fase G2 : 4-6 jam, replikasi
kromosom & distribusi ke
sel baru, G2/M check point
4. Fase M : 1 jam, Pembelahan
sel
KONTROL SIKLUS SEL (Fase G)
 REPLIKASI DNA (Fase S)

Replikasi DNA berlangsung pada kedua helix

dengan arah yang berlawanan
 REPLIKASI DNA (Fase S)
Garpu replikasi (replication fork ): leading strand and
lagging strand
Sintesis DNA baru : 5 3
Sintesis DNA leading strand 53 berlangsung
secara kontinu.
Sintesis DNA lagging strand tetap 53 tetapi
membentuk fragmen yang terpotong-potong yang
disebut Okazaki fragments
Okazaki fragments terdiri dari 100 200 nukleotida
DNA ligase menghubungkan Okazaki fragments,
sehingga membentuk untaian DNA utuh.
 Mitosis/Meiosis (Fase M)
Merupakan tahap pembelahan sel.
Interval waktu fase M kurang lebih 1 jam.
Pada mitosis, sel membelah dirinya
membentuk dua sel yang terpisah, sementara
pada meiosis sel membelah menjadi 4 sel.
Interfase : merupakan sebuah jedah panjang
antara satu mitosis dengan yang lain (Jedah
tersebut termasuk fase G1, S, G2).
Mitosis/Meiosis
(Fase M)
 BIOLOGI MOLEKULER ONKOLOGI

PROTOONKOGEN :
Gen yang normal memicu pembelahan sel yang
normal (proliferasi & diferensiasi), cth: ras, erb B,
src, bcl, myc, HER2.
ONKOGEN :
Protoonkogen yang bermutasi memicu pembelahan
sel yang tidak terkontrol (menjadi tumor)
TUMORSUPPRESSOR GEN :
Gen yang normal menghambat pembelahan sel.
Bila bermutasi akan menyebabkan pembelahan sel
menjadi tidak terkontrol, cth: Rb, p53, PTEN, BRCA.
 Produk Protoonkogen

SIS
ABL

FMS Inti Sel

SRC
Orga FOS
FMS
RAS nella MYC
JUN

MOS

ERB-B1
 ONCOGENE
Proto-oncogene Mutation Oncogene

Oncogenes :
promote cell proliferation
dominant & highly conserved
types:
viral oncogenes [v-oncs]
cellular oncogenes [c-oncs]
 ONCOGENE FAMILY

Classification of Oncogenes
A. Secreted Growth Factors
c-sis, hst
B. Cell Surface Receptors
erb B, fms, ret, trk, fes, fms Components of
signal transduction
C. Intracellular Transducers pathways
c-src, c-abl, mst, ras
D. DNA-binding Nuclear Proteins
myc, jun, fos
E. Regulators of the Cell Cycle
bcl, bax, bad
Molecular Biologi of Cancer. Burgers Medicinal Chemistry and Drug Discovery Sixth Edition, Volume 5:
Chemotherapeutic Agents. Edited by Donald J. Abraham. ISBN 0-471-37031-2 2003 John Wiley&Sons, Inc.
DOGMA SENTRAL
 SIGNAL
TRANSDUCTION
Cell Signaling
 TUMOR SUPPRESSOR GENE
TS Genes
inhibit growth and multiplication of mutated cells
prevent neoplastic transformation
recessive & highly conserved

Classification of TS genes
A. Cell Adhesion Molecules
APC, DCC
B. Regulators of the Cell Cycle
RB1, Tp53
 APOPTOSIS

APOPTOSIS programmed cell death

Balance between proliferation and apoptosis is

critical in determining growth or regression
Molecular Biologi of Cancer. Burgers Medicinal Chemistry and Drug Discovery Sixth Edition, Volume 5:
Chemotherapeutic Agents. Edited by Donald J. Abraham. ISBN 0-471-37031-2 2003 John Wiley&Sons, Inc.
 TUMOR SUPPRESSOR GENE

Retinoblastoma gene [RB1 gene]

rare form of childhood malignancy
forms: hereditary & sporadic
pRb
105-KDa nuclear protein
inhibits E2F [prevents G1 S transition]
inhibited by: phosphorylation viral
oncoproteins [E1A, HPV E7]
 TUMOR SUPPRESSOR GENE

p53 gene
location: 17p13.1
product: p53 protein [53 KDa]
function: induces DNA repair or apoptosis
mutation: point mutation > deletion
results to: loss of function p53
Clinical conditions: carcinomas, Li Fraumeni Syndrome
p53 inhibited by: HPV E6 (genomic), microRNA(epigenetic)
Faulty proteins can interfere with normal signal
transduction pathways

Path producing a product that inhibits cell division

Product of p53 tumor-suppressor gene is a
transcription factor

p53 transcription factor normally activates genes

for factors that stop DNA replication/ cell division

In the absence of functional p53, cell division

continues because the inhibitory protein is not
produced (p21)
 Cyclin Regulators
p 21: inhibits cell cycle progression and permits
DNA repair to take place.
P53: the guardian of the genome
In the presence of DNA damage, influences
transcription to either:
Halt cell cycle progression to facilitate DNA repair.
In cases of severe DNA damage, activates apoptosis.
Mutations in p53 are the most common genetic
alterations found in human cancer (50% of all human
tumors)
Growth Arrest

Video
Mechanisms of Oncogene Activation

1. Chrosomal Mutation
2. DNA Mutation
3. Viral gene integration
 1. Chromosomal Mutation
a. Chromosomal Deletion
Ex: Retinoblastoma
 1. Chromosomal Mutation
b. Chromosomal Amplification/ Duplication
Ex: Breast Ca, Cervical Ca, Lung Ca, Ovarian Ca, Hepatocellular
Ca, Esopagheal Ca, Colorectal Ca
 1. Chromosomal Mutation
c. Chromosomal Inversion
Ex: Papillary thyroid carcinomas
 1. Chromosomal Mutation
d. Chromosomal Translocation
d. Chromosomal Translocation

Ex. Burkitts Lymphoma

d. Chromosomal Translocation

Ex. Chronic Myelogenous Leukemia [CML]

 2. DNA MUTATIONS
A change in the DNA sequence of the gene

Mutations can alter protein product of DNA, stop

gene working or activate gene

Sudden, random alteration of original DNA code

that changes the genotype.

Can be caused by chemical or environmental

factors - mutagens.
 2. DNA MUTATIONS

Types of Mutation :

Deletion - DNA missing

Insertion - extra DNA inserted

Expansion - DNA repeat size has increased

Point Mutation - change in one base

 2. DNA MUTATIONS

Types of Mutation (in coding sequence)

AGC TTC GAC CCG Wild type (Normal)
AGC T_CG ACC CG Deletion
AGC TTC CGA CCC G Insertion
AGC TTC TTC GAC CCG Expansion
AGC TTC GAC CGG Point mutation
 POINT MUTATION

UAA
(Termination Codon)

UCA
(Codon for Serine)
UCU
(Codon for Serine)
CCA
(Codon for Proline)
QUESTION: What kind of mutation???
1. GTGCGGGCGATC
GTG CGG GCG ATC Wild type (Normal)
GTGCGGCCGATC
GTG CGG CCG ATC ???
POINT MUTATION

2. CCGTTCAGCGAC
CCG TTC AGC GAC Wild type (Normal)
CCGTCAGCGAC
CCG T_C AGC GAC ??? DELETION

AGC ATC GAC CGC

3. AGCATCGACCGC Wild type (Normal)
AGCATCGGACCGC
AGC ATC GGAC CGC ??? INSERTION
3. Viral Gene Integration

promoter

Viral promoter
 Viral Carcinogenesis
Viral carcinogens are classified into RNA and
DNA viruses.

Most RNA oncogenic viruses belong to the

family of retroviruses that contain reverse
transcriptase mediates transfer of viral RNA
into virus specific DNA.
 Viruses Associated With The
Development Of Human Neoplasia
VIRUSES NEOPLASMS
DNA VIRUSES
Human papilloma virus Cervical Ca, warts, ano-
genital carcinoma
Herpes simplex virus II Cervical carcinoma
Epstein-Barr virus NPCa, African Burkitts
Human Herpes virus 8 Kaposis sarcoma
Hepatitis B virus Hepatocellular Ca
Herpes simplex virus 6 Certain B cell
(HBLV) lymphomas
 Viruses Associated With The
Development Of Human Neoplasia

VIRUSES NEOPLASMS
RNA VIRUSES
Human T-
T-cell leukemia virus I Some T-T-cell leukemia,
lymphoma
Human T-
T-cell leukemia virus II Some cases of hairy
cell leukemia
Human immunodeficiency virus I Lymphoma; Kaposis
sarcoma
 VIRAL AGENTS: DNA viruses
Human Papillomavirus [HPV types 16, 18, 31, 33 & 35]

E2 is not expressed (E2: repression of E6/E7 genes)

Over-expression
E6 (inhibits p53)
E7 (inhibits pRb)
Transformasi sel pejamu akibat insersi sebuah promotor virus atau onkogen virus
CANCER CELLS
AND
NORMAL CELLS
Molecular Biologi of Cancer. Burgers Medicinal Chemistry and Drug Discovery Sixth Edition, Volume 5: Chemotherapeutic
Agents. Edited by Donald J. Abraham. ISBN 0-471-37031-2 2003 John Wiley&Sons, Inc.
Molecular Biologi of Cancer. Burgers Medicinal Chemistry and Drug Discovery Sixth Edition, Volume 5: Chemotherapeutic
Agents. Edited by Donald J. Abraham. ISBN 0-471-37031-2 2003 John Wiley&Sons, Inc.
INVASION-INTRAVASATION-METASTASIS
The defining characteristic of a malignancy.
Invasion: active translocation of neoplastic cells
across tissue barriers.
Critical pathologic point: local invasion and
neovascularization. These events may occur before
clinical detection.
 ANGIOGENESIS
Formation of new blood vessels from existing
vascular bed
Carried out by endothelial cells (EC) and extra
cellular matrix (ECM)
Regulated by angiogenic factors (inducers and
inhibitors)
* A tumor is unable to grow larger than 1 mm3
w/o developing a new blood supply
 ANGIOGENESIS
As tumor size increases, intratumoral O2 levels
fall and the center of the mass becomes
hypoxic leading to up-regulation of the
hypoxia inducible factor (HIF1)

An important transcriptional target of HIF1 is

the VEGF growth factor, induces
neovascularization of tumors
 INVASION-INTRAVASATION-METASTASIS

Mukherjee D. and Zhao J. 2013. The Role of chemokine receptor CXCR4 in breast cancer metastasis. Am J Cancer Res 3(1); 46-57.
 CARCINOGENS
Occupation related causes
Lifestyle related causes
Tobacco
Diet
Sexual practices
Viral carcinogens
Physical carcinogens
Chemical carcinogens
 Occupational Risk Factors

Etiology Site of Malignancy

Arsenic Lung, skin, liver
Asbestos Mesothelium, lung
Benzene Leukemia
Benzedine Bladder
Chromium cpds Lung
Radiation (mining) Numerous locations
Mustard gas Lung
Polycyclic hydrocarbons Lung, skin
Vinyl Chloride Angiosarcoma of liver
 Lifestyle Risk Factors
Tobacco-related:
Lung cancer
Pancreatic cancer
Bladder cancer
Renal cancer
Cervical cancer
 Diet-Related Risk Factors

Salt
Low vitamins A, C, E
Low consumption of Gastric Cancer
yellow-green Esophageal
vegetables Cancer
 Diet-Related Risk Factors

High fat Colon Cancer

Low fiber Pancreatic Cancer
Prostate Cancer
Low calcium
Breast Cancer
High fried foods
Uterine Cancer

Mycotoxins Liver Cancer

 Sexual Practices Risk Factors

Sexual promiscuity
Multiple partners
Unsafe Sex Cervical Cancer
Human Papillomavirus
 PHYSICAL CARCINOGENESIS

Radiation:
Ionizing - X-rays, rays, cosmic rays
Non-ionizing - UV light

Ionizing Radiation

includes electromagnetic rays & particulate matter

mechanism: free radicals & mutations
pathology: leukemia, thyroid ca, lung & breast ca
 RADIATION

PRE-IRRADIATION POST-IRRADIATION
 PHYSICAL CARCINOGENESIS

Ultraviolet Rays
UV-C filtered by ozone
UV-B

Inhibition of cell division

inactivation of enzymes
induction of mutations
cell death at high doses

Squamous cell cancer

Basal cell cancer
Melanocarcinoma
Efek radiasi atau karsinogen kimia pada protoonkogen atau promotor.
Mutasi dapat berupa point of mutation, delesi atau insersi
 DNA Mutation and Repair

Cells contain many DNA repair mechanisms.

Example - exposure to UV light.

UV - High energy light.

- Causes formation of thymine dimer.
- at low levels, it can be repaired.
- at high levels, it can be deadly.
 Photodimerization

Exposure to UV light UV light

can cause adjacent
thymines to
| | | | | | | |
covalently link. A C T T G C A T
T G A A C G T A
This results in a | | | | | | | |
distortion of the DNA
molecule and
breaks the hydrogen | | | | | |
thymine
A C G C A T
bonding with the dimer
T G C G T A
Adenine | | | | | |
 Nucleotide Excision Repair
(NER)
Separation
Operates by a cut-and patch mechanism that
removes a variety of bulky lesions, including
pyrimidines dimers and nucleotides which
Incision various chemical groups have become
attached.

Two pathways distinguished in

Excision NER
- Transcription-coupled pathway
(repairs the template)
- Global pathway (corrects DNA
Ligation
strands in the remainder of
the genome)
 Nucleotide Excision Repair (NER)

Damage recognition in the global pathway is mediated by an XPC protein

whereas damage recognition in the transcription-coupled pathways is thought
to be mediated by a stalled RNA polymerase in conjunction with a CSB
protein.

DNA strand separation (by XPB and XPD proteins, two helicase subunits of
TFIIH) (2)

Incision (by XPG on the 3side and the XPF-ERCC1 complex on the 5side) (3)

Excision (4)

DNA repair synthesis (by DNA polymerase and/or ) (5)

Ligation (by DNA ligase I).

 CHEMICAL CARCINOGENESIS

Carcinogens Promoters

cyclophosphamide saccharine & cyclamates

chlorambucil Estrogen
busulfan Diesthystilbestrol [DES]
melphalan
Base analogs
2-aminopurine
5-bromouracil
 IMUNOLOGI TUMOR

Sel tumor berbeda dengan sel normal dikenal sebagai

non-self/foreign (antigen)

Respon imun gagal menghambat pertumbuhan sel tumor

Antigen dikenal sebagai hasil :

Mutasi
Abnormal Expression
Oncogenic viruses
Oncofetal antigens
Altered surface modifications
Tissue specific differentiations
Olivera J. Finn, Ph.D. Cancer Immunology.N Engl J Med 2008;358:2704-15.
Tumor antigen, ada 2 tipe :
1. Tumor-Specific Antigens (TSA)/ Tumor-Specific
Transplantation Antigens (TSTAs)
Spesifik untuk tumor, tidak ada pada sel normal
Hasil mutasi pada sel tumor yang menghasilkan
perubahan pada protein seluler MHC kelas I induksi
respon imun dimediasi oleh CTL spesifik tumor
Protein yang dibentuk aibat mutasi 1 atau lebih gen
2. Tumor-Associated Antigens (TAAs)/ Tumor-
Associated Transplantation Antigens (TATAs)
Tidak spesifik untuk sel tumor
Hasil ekkspresi protein pada sel normal selama
pembentukan janin ( sistem imun masih belum matang
dan tidak dapat merespon)
Contoh : PSA, CEA, dll
Respon Imun Terhadap Tumor

Ada 4 jenis sel-sel imun yang berbeda dapat

membunuh target tumor in vitro maupun in vivo :

1. Sel pembunuh alami (Natural Killer / NK Cell)

2. Cytolytic thymus dependent Lymphocytes (CTLs)

3. Lymphokine-activated killer cells (LAK cells)

4. Macrophages
1. Natural Killer (NK) Cell
Sel-sel NK dapat membunuh sel-sel tumor tanpa
mensintesa sebelumnya antigen spesifik, aktivitasnya
tidak memerlukan adanya MHC kelas I pada sel-sel target.

NK Berkembang dalam bone marrow, kemudian

diperoleh dalam peripheral blood, sel pit (sinusoid liver)
dan sinusoid limpa

Dapat mensekresi interferon gamma, dan secara spontan

membunuh sel yang diinfeksi virus dan sel-sel tumor

Sel NK ambil bagian dalam pengawasan tumor yang mulai

timbul dan juga terhadap pertumbuhan metastatik
tumor.
Aktivasi Sel Natural Killer
2. CYTOLYTIC THYMUS-DEPENDENT LYMPHOCYTES
(CTLs) = Cytotoxic T cells

CTLs dapat membunuh tumor setelah dipresentasikan

oleh MHC kelas I

CTLs dapat membunuh sel-sel target melalui jalur yang

memerlukan cross-linked ligands CTLs dengan reseptor
pada permukaan spesifik sel-sel tumor untuk
merangsang apoptosis sel-sel tumor (program
kematian sel).
 AKTIVASI CYTOLYTIC THYMUS-DEPENDENT LYMPHOCYTES
(CTLs) = Cytotoxic T cells

Daniel S. Chen and Ira Mellman, Oncology Meets Immunology: The Cancer-Immunity Cycle. Immunity 39,
July 25, 2013. Elsevier Inc.
3. Lymphokine-Activated Killer cells
(LAK cells)

Subset null lymphocyte berbeda dari sel-sel

NK dan CTLs

Dapat dihasilkan in vitro, caranya mengkultur

sel-sel limfosit + Interleukin 2, IL2 konsentrasi
tinggi

Memiliki aktivitas anti tumor (membunuh sel

tumor).
Sistem Imun Tubuh
4. Macrophages
Sel fagosit mononukleus non limfosit
Ada pada jaringan dan dalam darah, derivat
dari stem sel monositic.
Penting sebagai sel pelengkap pada respon
imun
Makrofag khusus ada pada beberapa lokasi,
sel-sel Kupffer dan histiosit.
4. Macrophages
Diaktifkan sebagai suatu hasil dari reaksi delayed
Hypersentivity oleh sel T atau oleh aktivator makrofag
non spesifik, polinukleotida.

Peran: Sebagai scavengers , clean up injured cells

Dapat diaktifkan oleh berbagai agents, termasuk

peptida bakteri seperti: mycobacteria (BCG) dan
polinukleotida yang dianggap anntigen oleh tubuh

Mekanisme: membunuh dengan cara memfagosit,

melalui pembentukan oksigen radikal dan aktivasi
enzim proteolitik
Daniel S. Chen and Ira Mellman, Oncology Meets Immunology: The Cancer-Immunity Cycle. Immunity 39,
July 25, 2013. Elsevier Inc.
Daniel S. Chen and Ira Mellman, Oncology Meets Immunology: The Cancer-Immunity Cycle. Immunity 39,
July 25, 2013. Elsevier Inc.