Growth inhibitory action of Red Trapa natans shell methanolic extract (RTNSME) in Ehrlich ascites carcinoma (EAC) cells in Swiss Albino Mice by inducing apoptosis through inactivation of NFkB

Abstract Trapa natans L., commonly known as water chestnut, is a small aquatic herb belonging to the family Trapaceae. Its fruit is valued for both nutritional and pharmaceutical properties. This study investigates the anticancer potential of the methanolic extract of red Trapa natans shell (RTNSME) against Ehrlich ascites carcinoma (EAC) cells. RTNSME demonstrated significant anticancer activity, inhibiting EAC cell growth by 76.05% compared to the untreated control. Fluorescence microscopy of DAPI-stained cells revealed classic apoptotic features such as DNA fragmentation, nuclear condensation, cell shrinkage, and membrane alterations in RTNSME-treated cells. These findings were supported by RT-PCR analysis of apoptosis-related genes. RTNSME upregulated the expression of p53 and Bax, while downregulating the anti-apoptotic gene Bcl-2. Additionally, RTNSME suppressed the activity of the nuclear factor kappa B (NF-κB), further indicating its role in promoting apoptosis. GC-MS analysis of RTNSME identified 35 phytochemical constituents, including known bioactive and anticancer compounds such as canthaxanthin, dipyridamole, and colchiceinamide. Molecular docking studies suggested that canthaxanthin exhibits the strongest binding affinity and is likely the most potent compound against EAC cells. Overall, this study demonstrates that RTNSME exerts significant anticancer effects on EAC cells, primarily through NF-κB inactivation and induction of apoptosis.