Determination of the Effect of Androctonus crassicauda (Oliver, 1807) Crude Venom on Bcl-2 and Bax Genes Expression Levels in Colon Cancer Cell Line (HT-29) Using qPCR Method
Abstract
Introduction: Colorectal cancer (CRC) is one of the most common malignancies worldwide and remains a leading cause of cancer-related mortality, particularly when diagnosed at advanced stages. The search for novel anticancer agents derived from natural products has attracted increasing attention, with scorpion venoms representing a promising source of biologically active compounds. Material and Methods: This study investigated the in vitro cytotoxic effects of crude venom obtained from Androctonus crassicauda (Oliver, 1807) on human colorectal cancer cells. Human colorectal adenocarcinoma cells (HT-29) were used as the experimental model, whereas human embryonic kidney cells (HEK-293) served as the non-cancerous control. Results: Cell viability was assessed using the MTT assay, and the half-maximal inhibitory concentration (IC₅₀) values were determined to be 40.54 mg/mL for HT-29 cells and 57.30 mg/mL for HEK-293 cells. The expression levels of apoptosis-related genes were further analyzed by quantitative real-time PCR (qPCR). The analysis of gene expression levels using the qRT-PCR method showed that Bax gene expression decreased by approximately 0.25-fold in HT-29 cells compared with control group, whereas it increased approximately fivefold in HEK-293 cells. Similarly, Bcl-2 expression decreased to approximately 0.5-fold in HT-29 cells while increasing approximately 2.5- fold in HEK-293 cells. Conclusion: The lower IC₅₀ value observed in HT-29 cells indicates that Androctonus crassicauda crude venom exerts greater cytotoxic activity against colorectal cancer cells than against non-cancerous cells. Although the observed changes in apoptosis-related gene expression suggest that the venom modulates cellular death pathways, further studies are required to elucidate the underlying molecular mechanisms and to identify the bioactive venom components responsible for these effects. Overall, these findings support the potential of A. crassicauda venom as a promising natural source of bioactive molecules for the development of novel anti-cancer therapeutics. Keywords: Androctonus crassicauda, scorpion venom, colorectal cancer, cytotoxicity, apoptosis, HT-29, HEK-293, qPCR