Mitochondria-mediated anticancer effects of two previously characterized COX-2 inhibitors in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality and often shows limited response to current therapies like sorafenib. This study investigates the selective anticancer activity and mitochondria-mediated apoptotic effects of two previously characterized COX-2 inhibitors, ZMA-M3 (imidazo[1,2-a]pyrazine) and ZMA-M4 (imidazo[1,2-a]pyridine), in isolated HCC and normal rat hepatocytes. Isolated HCC and normal rat hepatocytes were treated with various concentrations of ZMA-M3 and ZMA-M4 to determine their half-maximal inhibitory concentrations (IC50). Apoptosis was evaluated using Annexin V-FITC/PI staining and caspase-3 activity assays. Mitochondrial dysfunction was assessed by measuring reactive oxygen species (ROS) generation, mitochondrial membrane potential (MMP) collapse, mitochondrial swelling, and cytochrome c release. Isolated HCC and normal rat hepatocytes were treated with various concentrations of ZMA-M3 and ZMA-M4 to determine their half-maximal inhibitory concentrations (IC50). Apoptosis was evaluated using Annexin V-FITC/PI staining and caspase-3 activity assays. Mitochondrial dysfunction was assessed by measuring reactive oxygen species (ROS) generation, mitochondrial membrane potential (MMP) collapse, mitochondrial swelling, and cytochrome c release.