This study aims to investigate the effect of ergosterol peroxide(EP)on the apoptosis of human hepatocellular carcinoma and its mechanism of action.The cell viability of HepG2 and SK-Hep-1 cells with 0(blank control),2.5,5,10,20,40,and 80 μmol·L-1 of EP after 24,48,and 72 h of action was detected by using CCK-8 assay,and the half inhibitory concentrations(IC50)at 24,48,and 72 h were calculated.Formal experiments were performed to detect the effect of EP on intracellular reactive oxygen species(ROS)using DCFH-DA staining,the effect of EP on intracellular mitochondrial membrane potential using JC-1 staining,the number of apoptotic cells using Annexin V-FITC/PI double-staining after HepG2 cells were co-cultured with 0(blank control),10,20,40 μmol·L-1 EP for 48 h.The effects of EP at different concentrations on apoptotic morphology were detected using AO/EB staining.The effects of different concentrations of EP on the protein expression of mitochondrial apoptosis pathway-related proteins B cell lymphoma 2(Bcl-2),cytochrome C(Cyt-C),Bcl-2-related X protein(Bax),caspase-3,cleaved caspase-3,caspase-9,and cleaved caspase-9 were examined by using Western blot.The results showed that different concentrations of EP could inhibit the proliferation of hepatocellular carcinoma with concentration-and time-dependent trends.Compared with the blank control group,the ROS level in the EP-treated group increased significantly(P<0.05).The mitochondrial membrane potential decreased significantly(P<0.05).The total apoptosis rate increased significantly(P<0.05).The expression of Bcl-2 protein was significantly down-regulated,and the expression of Cyt-C,Bax,cleaved caspase-9,and cleaved caspase-3 were significantly up-regulated(P<0.05).In summary,EP may inhibit the proliferation of hepatocellular carcinoma by modulating the mitochondria-mediated apoptosis pathway and induce apoptosis.
ergosterol peroxidehepatocellular carcinomamitochondrial membrane potentialmitochondrial apoptosisreactive ox-ygen species
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