AN IN SILICO AND IN VITRO EVALUATION OF CYTOTOXICITY, APOPTOTIC ACTIVITY AND GENE EXPRESSION MODULATION OF SARSASAPOGENIN IN HUMAN COLORECTAL CANCER CELL LINE HT-29

Abstract

Objective: Search for natural drugs against Colo Rectal Cancer (CRC) is ever-growing. Sarsasapogenin is a steroidal sapogenin known for various biological activities. The current study intends to investigate it’s anticancer activity in vitro against the Human Adenocarcinoma CRC cell line (HT-29). Additionally, the in silico interaction between sarsasapogenin and selected anticancer drug-protein targets was investigated. Methods: To evaluate cell viability, HT-29 cells were subjected to several concentrations of sarsasapogenin. Flow cytometry was used to study apoptosis. The expression of the genes Epidermal Growth Factor Receptor Tyrosine Kinase (EGFR-TK) and Kirsten Rat Sarcoma oncogene homolog (KRAS) was elucidated by real-time Polymerase Chain reaction. Molecular docking was used in conjunction with Molecular Dynamics (MD) simulation to comprehend the Sarsasapogenin’s interaction with EGFR-TK and KRAS. Results: Sarsasapogenin affected the viability of HT-29 cells dose-dependently. In HT-29 cells, sarsasapogenin treatment decreased the levels of KRAS and EGFR and caused apoptosis. In silico study demonstrated the interaction of sarsasapogenin in the Adenosine triphosphate binding site of EGFR-TK and the switch I/switch II site of KRAS. Post-MD analysis determined the stable binding of sarsasapogenin with these proteins. The binding energy with EGFR-TK and KRAS was found to be-46.0 ± 1.5 kcal/mol and-28.8 ± 6.3kcal/mol. Conclusion: Altogether, Sarsasapogenin, through modulation of EGFR and KRAS has shown promising anticancer effect against HT-29 cells.