Impact of a benzimidazole salt on gene expression, cytotoxicity, and apoptotic processes in HepG2 cell line

HepG2 is crucial in liver research because of its distinctive characteristics and significant applicability in drug metabolism, hepatotoxicity, and liver diseases. Benzimidazole derivatives are potential medicinal agents for the treatment of liver cancer. A new benzimidazole-based compound was created and verified using 1 H NMR, 13 C NMR, and IR. We assessed the effects of a novel benzimidazole salt compound 3 on HepG2, DLD-1, MCF-7, and HEK-293T cells in culture. Compound 3 was administered to each of the three cancer cell lines for 72 hours, and the IC 50 values were recorded. The IR, ADMET, real-time PCR, and the MTT assay were used to examine the effects of 3 . The viabilities of MCF-7, HepG2, and DLD-1 cells after 72 hours were 22.41 µM, 25.14 µM, and 41.97 µM, respectively. In HepG2 cells, the expression levels of many molecules relevant to pathways were measured in terms of mRNA and protein. Compound 3 exhibited significant cytotoxicity, with IC 50 values of 25.14 µM for HepG2, 22.41 µM for MCF-7, and 41.97 µM for DLD-1 cells, demonstrating selectivity toward cancer cells. Real-time PCR revealed elevated expression levels of pro-apoptotic markers (BAX, CASPASE-3, and CASPASE-8) and anti-apoptotic marker BCL-2, suggesting the induction of both intrinsic and extrinsic apoptotic pathways. The ADMET analysis highlighted favorable pharmacokinetic properties, including blood-brain barrier permeability and high gastrointestinal absorption. The novel benzimidazole compound 3 demonstrated significant anticancer activity and apoptotic potential in vitro , particularly against HepG2 cells. Its favorable ADMET profile and mechanism of action suggest its potential as a therapeutic agent for liver cancer. Future studies should focus on in vivo validation and further optimization of its pharmacological properties.