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Effects of standardized Cannabis sativa extract and ionizing radiation in

melanoma cells in vitro

Article  in  Journal of Cancer Research and Therapeutics · January 2018

DOI: 10.4103/jcrt.JCRT_1394_16

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Original Article

Effects of standardized Cannabis sativa

extract and ionizing radiation in melanoma
cells in vitro Jamal Naderi,
Nasim Dana,
Shaghayegh
ABSTRACT
Haghjooy
Context: Melanoma causes the highest number of skin cancer‑related deaths worldwide. New treatment methods are essential for Javanmard,
the management of this life‑threatening disease. Alireza
Aims: In this study, we investigated the efficacy of a standardized Cannabis sativa extract alone or in combination with single radiation Amooheidari1,
dose (6 Gy) in B16F10 mouse melanoma cells in an extract dose‑dependent manner. Maryam Yahay,
Golnaz Vaseghi2
Materials and Methods: C. sativa extract at three concentrations (25, 12.5, and 6.25 µg/mL) alone for 72 h or in combination
with radiation (24 h incubation after the extract treatment + 48 h incubation after exposure to radiation) were evaluated for cell Department of
viability of melanoma cells using 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide assay. Cells were also treated with Physiology, Applied
6.25 µg/mL extract alone for 72 h before analyzing C. sativa‑induced cell death by flow cytometry. Physiology Research
Center, Cardiovascular
Results: Administration of the extract alone or alongside radiation substantially inhibited melanoma cell viability and proliferation in Research Institute,
the extract dose response‑dependent manner. The inhibition of melanoma cell viability was paralleled by an increase in necrosis but Isfahan University
not apoptosis when melanoma cells were treated with the extract alone. Radiation alone did not have any antiproliferative effects, of Medical Sciences,
and radiation also did not synergize antiproliferative effects of the extract when the extract and radiation were combined.
1
Department of
Radiology, Milad
Conclusion: Our data suggest that C. sativa extract may have significant health and physiological implications for the treatment Hospital, 2Department
of melanoma. The results of this study also indicate that B16F10 mouse melanoma cells are radioresistant. Taken together, these of Pharmacology,
findings may lead to the identification of new therapeutic strategy for the management of melanoma. Isfahan Cardiovascular
Research Center,
Cardiovascular
Research Institute,
KEY WORDS: Anticancer effects, cancer treatment, cannabinoids, melanoma, radiation
Isfahan University
of Medical Sciences,
Isfahan, Iran
INTRODUCTION extracts and cannabinoids also have been useful
For correspondence:
in the pain management [10,11] and treatment
Dr. Golnaz Vaseghi,
“Cannabinoids” is a broad term covering a of inflammation. [11,12] The antiproliferative Department of
group of compounds extracted from the effects of cannabinoids in a variety of cancer Pharmacology, Isfahan
Cannabis sativa [1] and Cannabis indica plants. cell lines, including pancreatic, melanoma, Cardiovascular
D9‑tetrahydrocannabinol (THC) is the most glioma, lymphoma, lung, and breast, have been Research Center,
Cardiovascular
prominent active component in this group of reported.[3,13‑15] However, potential therapeutic
Research Institute,
various compounds. THC is mainly responsible interactions of Cannabis extracts and cannabinoids Isfahan University
for the psychoactive effects of the plant. [2,3] with conventional cancer therapies, such as of Medical Sciences,
Cannabinoids exert a variety of effects through radiation, are largely unknown. Exposure to Isfahan, Iran.
binding to specific receptors. At least two types of radiation leads to either necrosis or apoptotic cell E‑mail: golnazvaseghi@
yahoo.com
cannabinoid receptors (CBs) have been characterized death. The ability of different agents to promote the
as CB1 and CB2.[4,5] cell death is of central importance when selecting
specific anticancer treatment.[16] This study was Access this article online
Many studies have reported the antiemetic Website: www.cancerjournal.net
activity of Cannabis extracts and cannabinoids This is an open access journal, and articles are distributed under the terms of the DOI: 10.4103/jcrt.JCRT_1394_16
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which
on chemotherapy‑associated emesis. [2,6‑8] The allows others to remix, tweak, and build upon the work non-commercially, as
PMID: ***
results of preclinical trials have shown that long as appropriate credit is given and the new creations are licensed under the Quick Response Code:
THC possesses the antiemetic activity in animal identical terms.
models in radiation‑induced emesis.[8,9] Cannabis For reprints contact: reprints@medknow.com

Cite this article as: Naderi J, Dana N, Javanmard SH, Amooheidari A, Yahay M, Vaseghi G. Effects of standardized Cannabis
sativa extract and ionizing radiation in melanoma cells in vitro. J Can Res Ther 0;0:0.

© 2018 Journal of Cancer Research and Therapeutics | Published by Wolters Kluwer - Medknow 1
[Downloaded free from http://www.cancerjournal.net on Saturday, April 28, 2018, IP: 176.102.238.143]
Naderi, et al.: Effects of C. sativa extract and radiation in melanoma
performed to find whether C. sativa extract treatment alone or
in combination with single radiation dose (6 Gy) may reduce
the cell viability paralleled by increased cell death in mouse
melanoma cells in vitro.
Melanoma is the third‑most common skin cancer but the
leading cause of skin cancer‑related deaths.[17] The incidence
of melanoma is increasing worldwide, and the prognosis for
patients with advanced disease remains weak.[18] Treatment
options for melanoma include surgical excision, chemotherapy,
radiation therapy (RT), immunotherapy, targeted therapy,
prevention of angiogenesis, and combination therapies.[19‑21]
Surgery is the initial treatment for primary and locoregional
melanoma, but the other treatment options above are applied
for metastatic setting.[20] Although melanoma has been known a
radio‑resistant tumor, emerging data have shown that RT is now
recognized an effective therapy in some settings.[20] In addition
to current treatments, medicinal plants are important for cancer
treatment.[22,23] Effects of medicinal plants in various cancer
cell lines[24,25] including melanoma cells have been studied.[26,27]
Although there are different treatment options for melanoma,
any single treatment remains largely unsuccessful due to
the poor duration of response, resistance, and side effects.[21]
Therefore, the aim of this study is to examine the response
of mouse melanoma cells in vitro following C. sativa extract
treatment alone or in combination with radiation. Cell viability
and induction of apoptosis/necrosis were assessed in this study.
MATERIALS AND METHODS
Plant extraction
The plant was identified at the Botany Department of
the Faculty of Sciences, Isfahan University, and a voucher
specimen of the plant has been deposited in the herbarium of
Faculty of Pharmacy, Isfahan University of Medical Sciences,
Isfahan, Iran. C. sativa dry flowers and leaves were extracted
at room temperature with CO2 and standardized based on
4% cannabidiol, according to the information provided by the
supplier company (Barij‑Esans Co., Iran).
Reagents
Dulbecco’s modified eagle medium (DMEM) media, fetal
bovine serum (FBS), and Trypsin were from Gibco‑BRL (UK).
3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium
bromide (MTT) was purchased from Sigma‑Aldrich (Germany).
Annexin V‑FITC Apoptosis Detection Kit was obtained from
eBioscience (Austria).
Cell culture
The B16F10 mouse melanoma cell line (obtained from Pasteur
Institute) was cultured in DMEM supplemented with 10%
FBS at 37°C in a humidified atmosphere containing 5% CO2.
Cells were cultivated until 3–4 days to reach nearly 80%
confluency. The extract of C. sativa was added to the cells once
in DMEM/10% FBS. Untreated cells without the C. sativa extract
were included for unstimulated control.
2 Journal of Cancer Research and Therapeutics - Volume XX - Issue XX - Month 2018
Radiation therapy
The single absorbed dose of ionizing radiation (IR) was 6 Gy
from 6 MV photons at a source‑to‑surface distance of 100 cm.
The IR was delivered by an RT machine (Siemens ONCOR linear
accelerator) at room temperature.
3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide
assay
Following almost 80% confluence, melanoma cells were plated
into two 96‑well plates (3000 cells/well) and allowed to adhere
overnight. The cells, in one plate, were then pretreated for
24 h with varying doses of C. sativa extract (25, 12.5, and
6.25  µg/mL), before being subjected to the single radiation
dose (6 Gy). The cells, in this plate, were incubated for 48 h
following exposure to IR. The cells, in another plate, only
were treated for 72 h with the same doses of the extract as
the first plate, without IR. Following these procedures, MTT
was added to these two plates according to the instruction of
the supplier company and incubated for 4 h. The absorbance
was measured by a microplate reader (BioTek, OD 570 nm with
630 nm correction). The experiments were performed in series
of 3 wells per treatment and repeated three times.
Flow cytometry
Following almost 80% confluence, cells were seeded into
24‑well plates (1,00,000 cells/well) and left to adhere overnight.
Cells were treated with the lowest effective dose of the
extract (6.25 µg/mL) for 72 h. Following 72 h of treatment,
cells were washed in PBS followed by resuspending cells
in Binding Buffer. Annexin V‑FITC was added to the cell
suspension and incubated for 10 min at room temperature.
Propidium iodide (PI) was added to cell suspension according
to the information provided by the supplier company. Samples
were analyzed using a BD FACSCalibur flow cytometer in
triplicate in one independent biological replicate.
Statistical analysis
All data are shown as mean ± standard deviation. Differences
between variable and untreated control groups were
determined by appropriate ANOVA followed by Tukey–Kramer
test when appropriate. Statistical analysis was performed
using SPSS Statistics version 21. P ≤ 0.05 was considered
statistically significant.
RESULTS
C. sativa extract reduces the cell viability in B16F10
melanoma cells
We performed MTT assays to determine if the C. sativa extract
alone or alongside radiation would affect cell metabolic activity
as decreased metabolic activity may represent inhibition
of cell proliferation or decreased cell viability. Actively
proliferating melanoma cells were treated with three doses of
C. sativa extract (25, 12.5, and 6.25 µg/mL) alone for 72 h or in
combination with radiation (24 h incubation after the extract
treatment + 48 h incubation after exposure to radiation). MTT
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Naderi, et al.: Effects of C. sativa extract and radiation in melanoma
was then added and metabolized for 4 h. 25, 12.5, and 6.25 µg/mL
C. sativa extract treatments significantly decreased the cell
metabolic activity by 93.45%, 91.73%, and 87.76%, respectively
relative to untreated cells (n = 3; P ≤ 0.05) [Figure 1]. 25, 12.5,
and 6.25 µg/mL C. sativa extract treatments in combination with
radiation significantly decreased the cell metabolic activity by
92.69%, 91.38%, and 86.62%, respectively, relative to untreated
cells (nonradiation untreated cells) (n = 3; P ≤ 0.05) [Figure 1].
There is no significant difference between the metabolic activity
of treated cells by C. sativa extract and C. sativa extract‑treated
cells + radiation [Figure 1]. Therefore, the combination
treatment of C. sativa extract + radiation did not produce a
synergistic reduction of the metabolic activity. Radiation alone
also did not have any significant effects on the metabolic
activity of melanoma cells.
Extract of C. sativa induces cell death
To determine the possibility that cell death might have
contributed to the reduction in the metabolic activity and
cell viability detected, the induction of apoptosis and/or
necrosis was analyzed by annexin V‑FITC and PI. We did not
examine the effect of radiation alongside the C. sativa extract
in melanoma cells since the combination treatment of the
C. sativa extract + radiation did not produce a synergistic
reduction in the cell viability, and radiation alone also did not
have any significant effects on the cell viability of melanoma
cells. The dose of 6.25 µg/mL C. sativa extract was selected
to investigate cell death as this dose has been the lowest
effective dose of the extract in this study, which has been
Figure 1: Assessment of the metabolic activity in response to the
Cannabis sativa extract alone or in combination with radiation
using 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide.
B16F10 cells were exposed to three doses of the extract alone for
72 h or in combination with 6 Gy radiation (24 h incubation after the
extract treatment + 48 h incubation after exposure to radiation). Data
reflect mean  ±  standard deviation where untreated control is set to
100% (n = 3). *P ≤ 0.05 versus untreated control
Journal of Cancer Research and Therapeutics - Volume XX - Issue XX - Month 2018
evaluated, and this dose reduced the cell viability less than
other doses (25 and 12.5 µg/mL). Therefore, cells were treated
with 6.25 µg/mL extract for 72 h, stained with annexin V
and PI before analyzing C. sativa‑induced cell death by flow
cytometry. The melanoma cells responded to the extract
treatment as the percentage of healthy, necrotic, and apoptotic
cells was 79.66%, 20.04%, and 0.30%, respectively, compared
with untreated cells that the percentage of their healthy,
necrotic, and apoptotic cells was 98.48%, 0.56%, and 0.96%,
respectively (n = 1) [Table 1]. Necrosis was clearly detected
in response to the extract treatment according to our results.
Taken together, this finding corresponds to the data from the
MTT assay, supporting that C. sativa extract promotes cell death
in mouse melanoma cells.
DISCUSSION
This study shows that B16F10 mouse melanoma cells respond
to C. sativa extract alone or alongside radiation (6 Gy) by
reducing the cell viability in a C. sativa extract dose‑dependent
manner. However, our in vitro data suggest that the mouse
melanoma cells are radioresistant. Flow cytometry data show
that C. sativa extract at 6.25 µg/mL for 72 h significantly
induces necrosis. This result supports the data from MTT assay,
which C. sativa extract reduces the cell viability paralleled by
an increase in the cell death.
Both marijuana tar extract and synthetic THC were found
to hinder apoptosis while stimulating necrosis.[28] Cannabis
leaves have been shown to promote necrotic cell death by
secreting THA acid (THCA) into leaf tissues. THCA treatment
also promotes necrotic cell death in plants which do not
produce cannabinoids.[29] Our results are in agreement with
the results specified above. We observed that C. sativa extract
at 6.25 µg/mL for 72 h significantly induces necrotic cell death
while inhibiting apoptosis in melanoma cells. Further study
is required to find whether the active components in C. sativa
or lower doses of C. sativa extract may induce apoptosis in
melanoma cells in vitro. It is important to highlight that THC
is not cytotoxic at concentrations <5 µg/mL in A549 lung
tumor cells, but induces necrosis at higher levels with a lethal
concentration 50 = 16–18 µg/mL.[30] The study demonstrated
that C. sativa extract is cytotoxic at three concentrations
(6.25, 12.5, and 25 µg/mL), investigated in the present study,
and the extract at 6.25 µg/mL also induces necrosis. This
inconsistency in our results with results of Sarafian et al.[30] may
arise from using C. sativa extract instead of THC, different cell
types (B16F10 melanoma cells and A549 lung tumor cells), and
Table 1: Effects of the Cannabis sativa extract (6.25 µg/ml)
on cell death in B16F10 melanoma cells following 72 of
treatment as shown by flow cytometry
Healthy Necrotic Apoptotic
Control 98.48 0.56 0.96
Treatment 79.66 20.04 0.30
Each data represent percentage of cells (%) (n=1)
3
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Naderi, et al.: Effects of C. sativa extract and radiation in melanoma
incubation time. It is noteworthy that cannabinoids, primarily
THC, have demonstrated psychotropic effects.[31] Hence,
further study is needed to examine whether lower doses of
the C. sativa extract can reduce cytotoxicity and psychotropic
effects of the extract in vitro and in vivo.
Several studies support a role for Cannabis extracts and
cannabinoids in growth inhibition of tumor cells or tumor
cell death.[32‑35] Our results are consistent with aforementioned
studies and show that C. sativa extract significantly reduces
cell viability of melanoma cells and induces cell death. Further
investigation is required to find whether these effects of
C. sativa extract are selective and whether C. sativa extract
may have antitumor effects in vivo as well.
Although melanoma has historically been considered a relative
radio‑resistant tumor, RT has been recommended in a number
of clinical settings including definitive, adjuvant, and palliative
management of melanoma.[20,36,37] However, our data support
that B16F10 mouse melanoma cells are radioresistant. Our
finding is consistent with at least three studies[38‑40] that
showed melanoma is intrinsically radio‑resistant in vitro. On
the other hand, the response of melanoma to radiation has
been shown to depend on tumor volume, radiation dose, and
fraction size.[41] In the present study, single‑radiation dose was
6 Gy while early in vitro studies of melanoma radiosensitivity
by clonogenic assays indicated that successful treatment
required high dose per fraction RT.[42,43] Theoretically, the ability
of melanoma cell to conquer sublethal DNA injuries induced
by radiation proposes that, clinically, melanoma should be
more sensitive to large doses per fraction than to lower
fraction doses.[41] This hypothesis is in contrast to at least two
randomized clinical trials, showing similar rates of response
with low–to‑medium dose fractions (2.5–5 Gy) and larger
dose fractions (8–9 Gy). Moreover, the clinical trial showing
an improvement in regional disease control using low dose
per fraction (2.4 Gy) RT, further dispelling the myth that RT
is only effective for melanoma if given with high doses per
fraction.[43] Although the study of Mahadevan et al.[20] proposes
that the fractionation in melanoma is controversial, the
evidence suggests that clinicians should be comfortable with
using low‑dose‑per‑fraction RT in proper settings.[43] It should
be taken into consideration that the rate of cell viability for
melanoma cells exposed to single radiation doses in vitro was
found to differ markedly among individual cell lines.[42,43] Thus,
melanoma is a tumor type that shows considerable variation in
radioresponsiveness.[42,44,45] Thus, optimal RT of melanoma will
probably requires a personalized treatment approach. In vitro
experiments for prediction of radiocurability, the optimum
utilization of RT, and choice of treatment option for individual
melanoma patient appear therefore clearly warranted.[42]
The single doses of gamma rays and protons in the range
from 8 to 24 Gy following 7 days after irradiation significantly
reduced estimated cell viability in HTB140 human melanoma
cells, as compared to nonirradiated control. [16] Although
4 Journal of Cancer Research and Therapeutics - Volume XX - Issue XX - Month 2018
it was initially presumed that apoptosis ensued relatively
fast (within hours) after radiation, the evidence demonstrates
that apoptosis can ensue at long times after radiation
(out to 20 days) in some cell types.[46] Our results may confirm
the aforementioned studies as in the present study melanoma
cells did not respond to radiation following 48 h postradiation
incubation. Further studies are needed to find whether
melanoma cells may respond to radiation following longer
postradiation incubation time.
The effect of adjuvant RT in the treatment of melanoma
remains controversial and is underused.[37,47] Our study is
the first research that investigated the combined adjuvant
cellular effects of C. sativa extract and radiation in mouse
melanoma cells in vitro. This in vitro study demonstrates that
radiation (6 Gy) does not have any synergistic antitumor
effects in melanoma when combined with C. sativa extract in
an extract dose‑dependent manner.
CONCLUSION
This study provides the first evidence of antitumor effects of
C. sativa extract, when administered alone or in combination
with radiation, to mouse melanoma cells in vitro. Our results
may verify the value of C. sativa extract for the treatment
of melanoma and may complement the therapeutic profile
of C. sativa extracts administration in the future. Precise
mechanisms and the biologic basis of C. sativa extract‑induced
cell death need to be explored with well‑designed studies.
Further studies are required to evaluate antitumor effects of
C. sativa in vivo and also to investigate in vitro cellular effects
of combined C. sativa extract and radiation in different single
and fractionated doses of radiation, and longer postradiation
incubation time.
Acknowledgment
We thank Dr. Abed, Barij‑Esans Co., for kindly providing the
extract.
Financial support and sponsorship
This study was funded by Isfahan University of Medical
Sciences (grant number 293212).
Conflicts of interest
There are no conflicts of interest.
REFERENCES
1. Di Marzo V. Cannabinoids. 1st ed. Hoboken, NJ, US: Wiley‑Blackwell;
2014.
2. Grotenhermen F, Russo E. Cannabis and Cannabinoids: Pharmacology,
Toxicology, and Therapeutic Potential. 1st ed. Abingdon, England:
Routledge; 2002.
3. Scott KA, Dalgleish AG, Liu WM. The combination of cannabidiol
and Δ9‑tetrahydrocannabinol enhances the anticancer effects of
radiation in an orthotopic murine glioma model. Mol Cancer Ther
2014;13:2955‑67.
[Downloaded free from http://www.cancerjournal.net on Saturday, April 28, 2018, IP: 176.102.238.143]
Naderi, et al.: Effects of C. sativa extract and radiation in melanoma
4. Soethoudt M, Grether U, Fingerle J, Grim TW, Fezza F, de Petrocellis L,
et al. Cannabinoid CB2 receptor ligand profiling reveals biased
signalling and off‑target activity. Nat Commun 2017;8:13958.
5. Pertwee RG, Howlett AC, Abood ME, Alexander SP, Di Marzo V,
Elphick MR, et al. International union of basic and clinical
pharmacology. LXXIX. Cannabinoid receptors and their ligands:
Beyond CB1 and CB2. Pharmacol Rev 2010;62:588‑631.
6. Stewart DJ. Cancer therapy, vomiting, and antiemetics. Can J Physiol
Pharmacol 1990;68:304‑13.
7. Marmor JB. Medical marijuana. West J Med 1998;168:540‑3.
8. Olver I, Molassiotis A, Aapro M, Herrstedt J, Grunberg S, Morrow G,
et al. Antiemetic research: Future directions. Support Care Cancer
2011;19 Suppl 1:S49‑55.
9. Darmani NA, Janoyan JJ, Crim J, Ramirez J. Receptor mechanism and
antiemetic activity of structurally‑diverse cannabinoids against
radiation‑induced emesis in the least shrew. Eur J Pharmacol
2007;563:187‑96.
10. Schröder S, Beckmann K, Franconi G, Meyer‑Hamme G, Friedemann T,
Greten HJ, et al. Can medical herbs stimulate regeneration or
neuroprotection and treat neuropathic pain in chemotherapy‑induced
peripheral neuropathy? Evid Based Complement Alternat Med
2013;2013:423713.
11. Abrams DI. Integrating cannabis into clinical cancer care. Curr Oncol
2016;23:S8‑14.
12. Zheng D, Bode AM, Zhao Q, Cho YY, Zhu F, Ma WY, et al. The cannabinoid
receptors are required for ultraviolet‑induced inflammation and skin
cancer development. Cancer Res 2008;68:3992‑8.
13. Velasco G, Sánchez C, Guzmán M. Towards the use of cannabinoids
as antitumour agents. Nat Rev Cancer 2012;12:436‑44.
14. Emery SM, Alotaibi MR, Tao Q, Selley DE, Lichtman AH, Gewirtz DA,
et al. Combined antiproliferative effects of the aminoalkylindole
WIN55,212‑2 and radiation in breast cancer cells. J Pharmacol Exp
Ther 2014;348:293‑302.
15. Haustein M, Ramer R, Linnebacher M, Manda K, Hinz B. Cannabinoids
increase lung cancer cell lysis by lymphokine‑activated killer cells
via upregulation of ICAM‑1. Biochem Pharmacol 2014;92:312‑25.
16. Ristic‑Fira AM, Todorovic DV, Koricanac LB, Petrovic IM,
Valastro LM, Cirrone PG, et al. Response of a human melanoma
cell line to low and high ionizing radiation. Ann N Y Acad Sci
2007;1095:165‑74.
17. Olszanski AJ. Current and future roles of targeted therapy and
immunotherapy in advanced melanoma. J Manag Care Spec Pharm
2014;20:346‑56.
18. Garbe C, Eigentler TK, Keilholz U, Hauschild A, Kirkwood JM.
Systematic review of medical treatment in melanoma: Current status
and future prospects. Oncologist 2011;16:5‑24.
19. Thompson JF, Scolyer RA, Kefford RF. Cutaneous melanoma. Lancet
2005;365:687‑701.
20. Mahadevan A, Patel VL, Dagoglu N. Radiation therapy in the
management of malignant melanoma. Oncology (Williston Park)
2015;29:743‑51.
21. Davey RJ, van der Westhuizen A, Bowden NA. Metastatic melanoma
treatment: Combining old and new therapies. Crit Rev Oncol Hematol
2016;98:242‑53.
22. Utsuyama M, Seidlar H, Kitagawa M, Hirokawa K. Immunological
restoration and anti‑tumor effect by japanese herbal medicine in
aged mice. Mech Ageing Dev 2001;122:341‑52.
23. Drasar P, Moravcova J. Recent advances in analysis of Chinese medical
plants and traditional medicines. J Chromatogr B Analyt Technol
Biomed Life Sci 2004;812:3‑21.
24. Lian Z, Niwa K, Gao J, Tagami K, Mori H, Tamaya T, et al. Association
of cellular apoptosis with anti‑tumor effects of the Chinese herbal
complex in endocrine‑resistant cancer cell line. Cancer Detect Prev
2003;27:147‑54.
25. Manosroi A, Akazawa H, Akihisa T, Jantrawut P, Kitdamrongtham W,
Manosroi W, et al. In vitro anti‑proliferative activity on colon cancer
Journal of Cancer Research and Therapeutics - Volume XX - Issue XX - Month 2018
View publication stats
cell line (HT‑29) of Thai medicinal plants selected from Thai/Lanna
medicinal plant recipe database “MANOSROI III”. J Ethnopharmacol
2015;161:11‑7.
26. Carneiro Leite V, Ferreira Santos R, Chen Chen L, Andreu Guillo L.
Psoralen derivatives and longwave ultraviolet irradiation are active
in vitro against human melanoma cell line. J Photochem Photobiol B
2004;76:49‑53.
27. Finn GJ, Creaven BS, Egan DA. A study of the role of cell cycle events
mediating the action of coumarin derivatives in human malignant
melanoma cells. Cancer Lett 2004;214:43‑54.
28. Sarafian TA, Tashkin DP, Roth MD. Marijuana smoke and
delta (9)‑tetrahydrocannabinol promote necrotic cell death but inhibit
fas‑mediated apoptosis. Toxicol Appl Pharmacol 2001;174:264‑72.
29. Shoyama Y, Sugawa C, Tanaka H, Morimoto S. Cannabinoids act
as necrosis‑inducing factors in Cannabis sativa. Plant Signal Behav
2008;3:1111‑2.
30. Sarafian TA, Kouyoumjian S, Tashkin D, Roth MD. Synergistic
cytotoxicity of delta(9)‑tetrahydrocannabinol and butylated
hydroxyanisole. Toxicol Lett 2002;133:171‑9.
31. McAllister SD, Soroceanu L, Desprez PY. The antitumor activity of
plant‑derived non‑psychoactive cannabinoids. J Neuroimmune
Pharmacol 2015;10:255‑67.
32. Sarfaraz S, Adhami VM, Syed DN, Afaq F, Mukhtar H. Cannabinoids
for cancer treatment: Progress and promise. Cancer Res
2008;68:339‑42.
33. Scuderi MR, Cantarella G, Scollo M, Lempereur L, Palumbo M,
Saccani‑Jotti G, et al. The antimitogenic effect of the cannabinoid
receptor agonist WIN55212‑2 on human melanoma cells is mediated
by the membrane lipid raft. Cancer Lett 2011;310:240‑9.
34. Chakravarti B, Ravi J, Ganju RK. Cannabinoids as therapeutic agents
in cancer: Current status and future implications. Oncotarget
2014;5:5852‑72.
35. Romano B, Borrelli F, Pagano E, Cascio MG, Pertwee RG, Izzo AA,
et al. Inhibition of colon carcinogenesis by a standardized Cannabis
sativa extract with high content of cannabidiol. Phytomedicine
2014;21:631‑9.
36. Davar D, Tarhini AA, Kirkwood JM. Adjuvant therapy for melanoma.
Cancer J 2012;18:192‑202.
37. Shi W. Role for radiation therapy in melanoma. Surg Oncol Clin N
Am 2015;24:323‑35.
38. Doss LL, Memula N. The radioresponsiveness of melanoma. Int J
Radiat Oncol Biol Phys 1982;8:1131‑4.
39. Fertil B, Malaise EP. Intrinsic radiosensitivity of human cell lines is
correlated with radioresponsiveness of human tumors: Analysis
of 101 published survival curves. Int J Radiat Oncol Biol Phys
1985;11:1699‑707.
40. Petrović I, Ristić‑Fira A, Todorović D, Korićanac L, Valastro L, Cirrone P,
et al. Response of a radioresistant human melanoma cell line along
the proton spread‑out bragg peak. Int J Radiat Biol 2010;86:742‑51.
41. Strojan P. Role of radiotherapy in melanoma management. Radiol
Oncol 2010;44:1‑2.
42. Rofstad EK. Radiation biology of malignant melanoma. Acta Radiol
Oncol 1986;25:1‑0.
43. Barker CA. Radiation therapy for cutaneous melanoma:
Clonogenic assays to clinical trials. Oncology (Williston Park)
2015;29:752, 754.
44. Hill HZ, Hill GJ, Miller CF, Kwong F, Purdy J. Radiation and melanoma:
Response of B16 mouse tumor cells and clonal lines to in vitro
irradiation. Radiat Res 1979;80:259‑76.
45. Stevens G, McKay MJ. Dispelling the myths surrounding radiotherapy
for treatment of cutaneous melanoma. Lancet Oncol 2006;7:575‑83.
46. Held KD. Radiation‑induced apoptosis and its relationship to loss of
clonogenic survival. Apoptosis 1997;2:265‑82.
47. Khan N, Khan MK, Almasan A, Singh AD, Macklis R. The evolving role
of radiation therapy in the management of malignant melanoma.
Int J Radiat Oncol Biol Phys 2011;80:645‑54.
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