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目的研究昆明山海棠总生物碱(total alkaloids of tripterygium hypoglaucum hutch,THHta)对人结肠癌HCT116细胞增殖及凋亡的影响。方法采用MTT法检测THHta对HCT116细胞增殖的抑制作用;用倒置及荧光显微镜观察THHta作用后HCT116细胞形态的变化;流式细胞术检测THHta对HCT116细胞周期及凋亡的影响,Western bolt检测凋亡相关caspase-3及PARP蛋白表达。结果THHta能显著抑制HCT116细胞增殖,抑制率与剂量和时间呈依赖关系。THHta作用HCT116细胞48、72h的IC50值小于20μg/ml。THHta(10μg/ml)能诱导HCT116细胞G0/G1期比例增加及细胞凋亡,caspase-3活化及PARP蛋白剪切显著增强。结论THHta能显著抑制HCT116细胞增殖,并可诱导HCT116细胞G0/G1期阻滞及细胞凋亡。
Objective To investigate the effects of total alkaloids of tripterygium hypoglaucum hutch (THHta) on the proliferation and apoptosis of human colon cancer HCT116 cells. Methods The inhibitory effect of THHta on the proliferation of HCT116 cells was detected by MTT assay; the morphology changes of HCT116 cells after THHta treatment were observed by inversion and fluorescence microscope; the effect of THHta on cell cycle and apoptosis of HCT116 cells was detected by flow cytometry; apoptosis was detected by Western bolt. Related caspase-3 and PARP protein expression. Results THHta could significantly inhibit the proliferation of HCT116 cells. The inhibition rate was dependent on the dose and time. The IC50 values of THHta in HCT116 cells at 48 and 72 h were less than 20 μg/ml. THHta (10μg/ml) could induce the increase of G0/G1 phase ratio and cell apoptosis in HCT116 cells, and the activation of caspase-3 and the cleavage of PARP protein were significantly enhanced. Conclusion THHta can significantly inhibit the proliferation of HCT116 cells and induce G0/G1 phase arrest and apoptosis in HCT116 cells.