Snake venom includes various elements and therapeutically relevant nontoxic components, and a large number of pharmacologically beneficial compounds have been isolated and described. The purpose of this study was to compare the effects of native and gamma-irradiated Cerastes cerastes venom on breast & colon cancer cell lines. The anti-proliferative impact of the native and gamma-irradiated venoms determined using 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. Quantitative real time PCR used to evaluate the expression of apoptosis-related genes. Biochemical analysis Malondialdehyde (MDA) measured by the thiobarbituric acid assay, reduced Glutathione (GSH) was measured photometrically & ROS was assessed by 2, 7- dichlorofluorescein diacetate. Analytical SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Ultraviolet absorption Spectrum 200-900 nm. IC50 of gamma-irradiated venom was significantly reduced than native venom recorded half maximal inhibitory concentration (IC50) values of (19.93 ± 0.867 & 2.131 ± 0.095) μg/ml and (8.921 ± 0.515 & 0.9454 ± 0.055) μg/ml against human breast cancer (MCF-7) and human colon cancer (HCT-116) cell lines, respectively. Both venoms significantly induced the down-regulated Bcl-2 gene and up-regulated P53 and Bax genes. Both venoms significantly enhanced the antioxidant enzymes' activity levels of H2O2, while insignificantly reduced GSH enzyme activity. MDA was significantly elevated post treatment of native venom compared to irradiated venom and was a negative control for both cell lines. Gamma radiation can create changes in the components of snake venom, allowing it to be detoxified; gamma radiation has a significant effect on decreasing venom toxicity while having no influence on immunogenicity in the examined cell lines.