熊胆粉通过线粒体依赖性途径诱导人肝癌细胞凋亡

Objective:To evaluate the effect of Bear Bile Powder(熊胆粉,BBP) on the growth and apoptosis of HepG2 human hepatocellular carcinoma cells,and investigate the possible molecular mechanisms mediating its anti-cancer activity.Methods:HepG2 cells were treated with 0.4-1.0 mg/mL of BBP for 24,48 and 72 h.The viability of HePG2 cells was determined by MTT assay.Cellular morphology was observed via phase-contrast microscopy.Fluorescence-activated cell sorting analysis with Annexin-V/propidium idodide and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-benzimidazol-carbocyanine iodide(JC-1) staining was performed to determine cell apoptosis and the loss of mitochondrial membrane potential,respectively.Activation of caspase-9 and-3 was evaluated by a colorimetric assay.Results:The treatment with 0.4-1 mg/mL of BBP for 24,48,or 72 h respectively reduced cell viability significantly by 7%-60%,20%-90%or 25%-98%,compared with the untreated control cells(P0.01).In addition,BBP treatment induced morphological changes in HepG2 cells.Furthermore,after treated with 0,0.4,0.6,0.8 and 1.0 mg/mL of BBP,apoptosis cells(including early and late apoptotic cells) were 18.0%±1.3%,34.9%±2.2%,33.9%±2.8%,37.4%±2.8%and 46.0%±2.5%,respectively(P0.05);and the percentage of cells with reduced JC-1 red fluorescence were 6.6%±0.8%,8.5%±0.8%,13.5%±1.6%,17.6%±2.3%and46.7%±3.6%,respectively(P0.01).Finally,BBP treatment significantly and dose-dependently induced activation of both caspase-9 and caspase-3 in HepG2 cells(P0.05).Conclusions:BBP could inhibit the growth of HepG2hepatocellular cancer cells through mitochondrion-mediated apoptosis,which may,in part,explain its anti-cancer activity.BBP may be a potential novel therapeutic agent for the treatment of hepatocellular carcinoma.