Investigation of cytotoxic, anti-angiogenic and apoptotic effects of venom of snakes belonging to Vipera anatolica anatolica and Vipera anatolica senliki species in Turkey

Snake venoms are mixtures of bioactive proteins and peptides with various biochemical activities. Venomous snakes commonly found in our country are members of the Viperidae family. The protein contents of some species belonging to this family have been investigated by electrophoretic methods, but detailed studies on their mechanism of action have not been carried out. Snake venoms represent a source of potentially valuable therapeutic compounds due to the potent biological activity of the toxins they contain. In this thesis thesis, the cytotoxic, anti-angiogenic and apoptotic effects of Vipera anatolica anatolica (VAA) and Vipera anatolica senliki (VAS) venoms belonging to the Vipera anatolica species, were investigated. Venoms belonging to VAA and VAS species were collected in field conditions in Antalya province and its surroundings and lyophilized in Eppendorf tubes at +4℃ to the laboratory environment. Protein concentration determination was made by the Bradford method. The MTT method was used to determine the cytotoxic effect. The concentration (IC50) values that killed 50% of the cells were calculated using the CalcuSyn program. Apoptotic cells were determined in flow cytometry using the Annexin V Apoptosis Detection Kit. The scratch assay method, which detects the amount of cell migration, was used to investigate the anti-angiogenic effect. Protein amounts of VAA and VAS poisons were determined as 124.6±0.4 mg/mL and 141.8±1.2 mg/mL, respectively. It was determined that both snake venoms showed cytotoxic effects in both cancer cells and non-tumorigenic cells depending on concentration and time (p<0.05). The IC50 values of VAA venom in PC-3, MCF-7, H69 and HEK-293 cells were determined as 4.0 ± 0.8, 4.1 ± 1.2, 6.3 ± 0.6 and 6.0 ± 1.1, respectively. The IC50 values of VAS venom in PC-3, MCF-7, H69 and HEK-293 cells were determined as 5.8 ± 0.2, 5.2 ± 0.7, 6.2 ± 1.0 and 8.1 ± 1.2, respectively. With the IC50 value of VAA venom, PC-3 cells treated for 48 hours were 22.7% early apoptotic, 17.9% late apoptotic cells, MCF-7 cells 50.6% early apoptotic, 33.1% late apoptotic cells, H69 cells 25.7% early apoptotic, 66.2% late apoptotic cell rate was determined. The IC50 value of VAS venom and the rate of 8.0% early apoptotic 20.9% late apoptotic cells in PC-3 cells treated for 48 hours, 40.8% early apoptotic 19.5% late apoptotic cells in MCF-7 cells, 25.7% early apoptotic 66.2% late apoptotic cells in H69 cells determined. (p<0.05). Wound repair results revealed that VAA and VAS poisons inhibited the migration of cancer cells at 48 hours. With this thesis, it was shown for the first time that venoms obtained from VAA and VAS have cytotoxic effects and that their cytotoxic effects induce apoptotic cell death, as well as have potential anti-angiogenic effects. It is anticipated that the preliminary data to be obtained from snake venoms may enable the discovery of new pharmaceutical peptides and the discovery of more effective treatment methods for cancer or other diseases. With the current thesis, it has been shown that VAA and VAS poisons have anti-cancer effects, and these effects need to be confirmed by in vivo methods.