Mechanism of inducing mitophagy in human hepatocellular carcinoma Hep3B cells by Bakuchiol
[Objective]To investigate the effect of bakuchiol(BAK)on the mitophagy of human hepatocellular carcinoma Hep3B cells and its Mechanism.[Methods]MTT assay was used to detect the effect of BAK and mitophagy inhibitors CQ/Baf-A1 on the proliferation of Hep3B cells;Morphological changes of Hep3B cells after BAK treatment observed by phase contrast microscopy;Hoechst/Mitotracker/Rhodamine fluorescent probe staining combined with high connotation cell imaging system was used to analyze the effect of BAK on mitochondrial function;The expression of autophagy-related proteins LAMP1,LC3,p62,and mitophagy-related proteins PINK1,Parkin,MFN2,Tom20 at different time points of BAK were detected by Western blotting.[Results]BAK inhibited the growth of Hep3B cells in the time-and dose-dependent manners;High Content Analysis results suggest that BAK can elevate mitochondrial mass and decrease mitochondrial membrane potential;The addition of mitophagy inhibitors markedly reverse the death rate of Hep3B cells induced by BAK;The proteins LAMP1,LC3,PINK1,Parkin were significantly increased by BAK in the time-dependent manners,while the p62,MFN2,Tom20 were reduced;Addition of mitochondrial autophagy inhibitor could significantly reverse the growth inhibition of Hep3B cells induced by BAK.[Conclusion]Bakuchiol induces inhibition of cell growth in Hep3B cells by activating mitophagy.
国家科技期刊平台
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