AdipoRon Promotes Mitochondrial Ca^(2+)Overload and Apoptosis in Hepatocellular Carcinoma Cells by Activating the PLC-IP3-IP3R Signaling PathwayOA

Objective:Hepatocellular carcinoma(HCC)ranks among themost prevalentmalignant tumors globally.Metabolically associated fatty liver disease is a significant risk factor for HCC.Adiponectin,a key regulatory protein in glucolipid metabolism,presents potential as an anti-tumor target in HCC cells.The study focused on evaluating the anti-HCC properties of AdipoRon,an agonist of the adiponectin receptor.Method:Cell viability and proliferation were assessed using the cell counting kit-8 and colony formation assays,respectively.AdipoRon’s effect on HCC cell damage was evaluated via flow cytometry,apoptosis,and(lactate dehydrogenase)LDH assays.Mitochondrial function was evaluated by measuring mitochondrial membrane potential(MMP),ATP levels,and Complex I activity.Additionally,mitochondrial reactive oxygen species(ROS)and calcium(Ca^(2+))levelswere analyzed usingMitoSOXRed and Rhod-2 AM probes,respectively.Results:Our findings indicated that AdipoRon suppressed the proliferation of HCC cells and triggered apoptosis,with both effects being dose-dependent.Furthermore,AdipoRon caused a decrease in mitochondrial membrane potential,ATP levels,and Complex I activity,alongside the generation of mitochondrial ROS.Notably,AdipoRon disrupted intracellular Ca^(2+)homeostasis by causing mitochondrial Ca^(2+)overload due to release fromthe endoplasmic reticulum(ER).Additionally,AdipoRon promoted Ca^(2+)release from the ER by activating the PLC-IP3-IP3R pathway.The resulting mitochondrial Ca^(2+)overload enhances the anti-HCC effect when combined with chemotherapeutic drugs.Conclusions:Therefore,our study demonstrates thatAdipoRon promotesmitochondrial Ca^(2+)overload and apoptosis in HCC cells by activating the PLC-IP3-IP3R signaling pathway.AdipoRon has the potential to become an effective anti-HCC drug.