Cardionogen-1, a novel small molecule, induces cytotoxicity by inhibiting Wnt/β-catenin signalling pathway in Huh-7 cells.

This study investigated the cytotoxic property of cardionogen-1 (CDNG-1), a small molecule Wnt/β-catenin signalling inhibitor in the human hepatoma cell line. Huh-7 cells were treated with 5, 10, and 20 µM/ml of CDNG-1 for 24 h. Cytotoxicity assay, intracellular reactive oxygen species (ROS) analysis, apoptotic evaluation, scratch assay, flow cytometry, PCR and western blotting were performed to investigate the anticancer effect of CDNG-1 in Huh-7 cells. CDNG-1 markedly inhibited the viability, proliferation, and migration of Huh-7 cells in a dose-dependent manner. Fluorescence microscopic analyses revealed typical characteristics of ROS-mediated early apoptosis in Huh-7 cells upon CDNG-1 treatment. CDNG-1 effectively induced intrinsic apoptosis in Huh-7 cells, evidenced by increased protein levels of p53, p21, Bax, and cytochrome c, and decreased protein levels of the anti-apoptotic Bcl-2. CDNG-1 disrupted the cell cycle and caused accumulation of Huh-7 cells at the G0/G1, S and G2/M phases. CDNG-1 was also found to reduce the protein expression of cell cycle regulators like cyclin D and CDK-4. Moreover, CDNG-1 inhibited the Wnt/β-catenin pathway, evidenced by downregulated gene expression of Wnt3, LRP6 and β-catenin. Therefore, this study suggests that CDNG-1 exerts potential anti-proliferative, anti-migratory, and pro-apoptotic effects in liver cancer cells in a concentration-dependent manner, and hence, it could serve as a promising anticancer candidate for targeted therapies against liver cancer.