الفهرس | Only 14 pages are availabe for public view |
Abstract Squamous cell carcinoma is the most common type of cancers in head and neck region. Over the last three decades, the survival rate for all type of cancers has increased significantly due to the successful development of targeted therapy. So, there is a necessity to continuously develop some new biologics which could work alone or in combinations to fight against the complex cancer disease. Treatment of cancer involves different clinical protocols including: surgery, radiotherapy, chemotherapy and gene therapy. It has been found that chemotherapy provides a significant improvement in organ preservation and a decrease in the rate of cancer metastasis. Recently widespread attention has been given to develop safe and effective anticancer agents from natural sources. In this study, we investigated the effect of Egyptian cobra (N. haje) venom on HEp-2 cell line after 24 hours. In our study, Dose-dependent cellular toxicity caused by N. haje venom on HEp-2 cells was demonstrated by five independent experiments: cell viability assay, microscopic examination, image morphometric analysis, FCM and Annexin-V/PI assays. These experiments revealed that the percentages of apoptotic and necrotic cells were significantly increased in dose dependent manner. Results of the present study showed that N. haje venom had a cytotoxic effect on HEp-2 cell line after 24 hours in comparison to control cells. Histopathologically, the current study revealed that, the major criteria of apoptotic cell death as well as necrotic cell death increased with increasing N. haje venom concentrations. However, apoptosis played the major part. On evaluation of the morphometric parameters, our data revealed a decrease in the mean NAF values of all treated HEp-2 cells in relation to control cells after 24 hours. NAF is considered as a good indicator of apoptotic cell death. Statistically, our data showed a significant decrease in the mean NAF values of HEp-2 treated cells when compared with control cells after 24 hours P value = 0.0001. Regarding to FCM, data obtained showed that N. haje venom produced pre-G1 apoptosis with cell cycle arrest at G2/M Phases. The percentages of cells in Pre-G1 and G2/M phases increased after treatment with N. haje venom when compared with control cells after 24 hours. Annexin-V/PI staining assays showed that the number of apoptotic and necrotic cells increased with increasing the dose of N. haje venom from pre IC50 then IC50 to post IC50. These results demonstrated that there was a positive correlation between decreased viability of HEp-2 cells and increased N. haje venom concentrations. |