昆明山海棠总生物碱对人结肠癌HCT116细胞增殖和凋亡的影响

Effects of total alkaloids of Tripterygium hypoglaucum Hutch on proliferation and apoptosis of human colon cancer cell line HCT116

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 cell line HCT116.Methods The effects of THHta on the proliferation of HCT116 cells were assessed by MTT assay.Morphological changes of HCT116 cells were observed with invert microscopy and fluorescence microscopy.The cell cycle distribution and apoptosis of HCT116 cells were examined by flow cytometry（FCM）.The expression of active caspase-3 and cleaved PARP were detected by Western blot assay. Results THHta inhibited the proliferation of HCTll6 cells in a concentration- and time-dependent manner. The IC50 values of THHta which affected HCT116 cells for 48 and 72 h were less than 20 μg/ml. THHta （10μg/ml） increased the percentage of cells at the G0/G1 phase, induced the apoptosis of HCTll6 cells, and increased the amount of active caspase-3 and cleaved PARP in HCT116 cells. Conclusion THHta inhibits proliferation, causes G0/G1 phase arrest and induces apoptosis in cer HCT116 cells in vitro.