Anti-proliferative and cytotoxic effect of Iranian snake (Vipera raddei kurdistanica) venom on human breast cancer cells via reactive oxygen species-mediated apoptosis

Ehsan Malekara , Mona Pazhouhi, Iraj Rashidi, Cyrus Jalili



Background and purpose: Breast cancer is the most commonly occurring cancer in women around                  the world. Despite new advances in cancer therapy, breast cancer remains a disease with high morbidity               and mortality. Snake venom is a poisonous mixture of different molecules, such as carbohydrates, nucleosides, amino acids, lipids, proteins, and peptides. Previous studies demonstrated that some snake venoms showed in vitro anti-cancer effects. In this study, the effects of the Iranian snake                                 (Vipera raddei kurdistanica) venom on breast cancer cells were investigated.

Experimental approach: The effect of increasing concentrations of snake venom on breast cell viability was assessed by trypan blue, MTT, and lactate dehydrogenase measurements. Apoptosis was detected                  and quantified by fluorescent staining and DNA fragmentation assay. The expression level of                            some apoptotic-related genes was investigated using real-time polymerase chain reaction (RT-PCR).                   The Western blotting method was also used to detect the protein expression profiles in the cells.

Findings / Results: After treatment for 24, 48, 72, and 96 h, the cell viability was significantly reduced                     in a time- and dose-dependent manner (P < 0.05). The venom effect on normal breast cells was significantly smaller than cancer cells (P > 0.05). Apoptosis was significantly increased (P < 0.05). The RT-PCR                     and western blot data confirmed the increase of apoptosis in cells treated with venom.

Conclusion and implications: These data suggested that the vipera raddei kurdistanica venom had                            a cytotoxic property via activation of apoptosis in breast cancer cells.

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Nagai H, Kim YH. Cancer prevention from the perspective of global cancer burden patterns. J Thorac Dis. 2017;9(3):448-451.

Sun YS, Zhao Z, Yang ZN, Xu F, Lu HJ, Zhu ZY, et al. Risk factors and preventions of breast cancer. Int J Biol Sci. 2017;13(11):1387-1397.

Power EJ, Chin ML, Haq MM. Breast cancer incidence and risk reduction in the Hispanic population. Cureus. 2018;10(2):1-12.

Shahali A, Ghanadian M, Jafari SM, Aghaei M. Mitochondrial and caspase pathways are involved in the induction of apoptosis by nardosinen in MCF-7 breast cancer cell line. Res Pharm Sci. 2018;13(1):12-21.

Gewirtz DA, Bristol ML, Yalowich JC. Toxicity issues in cancer drug development. Curr Opin Investig Drugs. 2010;11(6):612-614.

Housman G, Byler S, Heerboth S, Lapinska K, Longacre M, Snyder N, et al. Drug resistance in cancer: an overview. Cancers (Basel). 2014;6(3):1769-1792.

Shanbhag VKL. Applications of snake venoms in treatment of cancer. Asian Pac J Trop Biomed. 2015;5(4):275-276.

Li L, Huang J, Lin Y. Snake venoms in cancer therapy: past, present and future. Toxins (Basel). 2018;10(9):1-8.

Nalbantsoy A, Hempel BF, Petras D, Heiss P, Göçmen B, Iğci N, et al. Combined venom profiling and cytotoxicity screening of the Radde's mountain viper (Montivipera raddei) and Mount bulgar viper (Montivipera bulgardaghica) with potent cytotoxicity against human A549 lung carcinoma cells. Toxicon. 2017;135:71-83.

Khazaei M, Pazhouhi M. Antiproliferative effect of Trifolium pratens L. extract in human breast cancer cells. Nutr cancer. 2019;71(1):128-140.

Strober W. Trypan blue exclusion test of cell viability. Curr Protoc Immunol. Appendix 3: Appendix 3B. DOI: 10.1002/0471142735.ima03bs21.

Mohamadi A, Aghaei M, Panjehpour M. Estrogen stimulates adenosine receptor expression subtypes in human breast cancer MCF-7 cell line. Res Pharm Sci. 2018;13(1):57-64.

Kumar P, Nagarajan A, Uchil PD. Analysis of cell viability by the lactate dehydrogenase assay. Cold Spring Harb Protoc. 2018;2018(6). DOI: 10.1101/pdb.prot095497.

Liu K, Liu PC, Liu R, Wu X. Dual AO/EB staining to detect apoptosis in osteosarcoma cells compared with flow cytometry. Med Sci Monit Basic Res. 2015;21:15-20.

Khazaei M, Pazhouhi M, Khazaei S. Evaluation of hydro-alcoholic extract of Trifolium pratens L. for its anti-cancer potential on U87MG cell line. Cell J. 2018;20(3):412-421.

Khazaei M, Pazhouhi M. Temozolomide-mediated apoptotic death is improved by thymoquinone in U87MG cell line. Cancer Invest. 2017;35(4): 225-236.

Khazaei M, Pazhouhi M, Khazaei S. Temozolomide and tranilast synergistic antiproliferative effect on human glioblastoma multiforme cell line (U87MG). Med J Islam Repub Iran. 2019;33:39-46.

Mahmood T, Yang PC. Western blot: technique, theory, and trouble shooting. N Am J Med Sci. 2012;4(9):429-434.

Pan Y, Ma S, Cao K, Zhou S, Zhao A, Li M, et al. Therapeutic approaches targeting cancer stem cells. J Cancer Res Ther. 2018;14(7):1469-1475.

Pal SK, Gomes A, Dasgupta SC, Gomes A. Snake venom as therapeutic agents: from toxin to drug development. Indian J Exp Biol. 2002;40(12):1353-1358.

Mohammad RM, Muqbil I, Lowe L, Yedjou C, Hsu HY, Lin LT, et al. Broad targeting of resistance to apoptosis in cancer. Semin Cancer Biol. 2015;35:S78-S103.

Shi MD, Shiao CK, Lee YC, Shih YW. Apigenin, a dietary flavonoid, inhibits proliferation of human bladder cancer T-24 cells via blocking cell cycle progression and inducing apoptosis. Cancer Cell Int. 2015;15:33-44.

Juan ME, Wenzel U, Daniel H, Planas JM. Resveratrol induces apoptosis through ROS-dependent mitochondria pathway in HT-29 human colorectal carcinoma cells. J Agric Food Chem. 2008;56:4813-4818.


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