摘要
目的观察虎杖白藜芦醇对人结肠癌HCT-15细胞增殖及凋亡的影响。方法以0、25、50、100、200μmol/L白藜芦醇作用人结肠癌HCT-15细胞24h以及50μmol/L白藜芦醇处理HCT-15细胞0、12、24、48、72h。采用噻唑蓝(MTT)法检测白藜芦醇对HCT-15细胞增殖的影响,倒置相差显微镜观察细胞形态变化,流式细胞术检测细胞周期的分布以及细胞凋亡率,免疫印迹方法以及聚合酶链反应法观察主要的细胞周期蛋白以及基因的表达情况。结果虎杖白藜芦醇能显著抑制HCT-15细胞生长,并呈现时间-浓度依赖性。细胞形态学检测显示随着药物浓度升高,细胞明显皱缩。流式细胞检测显示,随着白藜芦醇浓度的增加以及作用时间的延长,G0/G1期细胞比例明显增加,其中0、50、100、200μmol/L白藜芦醇处理24h后分别为(42.81±1.21)%、(49.25±0.92)%、(54.67±1.03)%、(64.24±1.17)%,50μmol/L白藜芦醇作用0、24、48、72h后分别为(32.41±1.09)%,(48.35±1.03)%,(55.61±1.33)%,(70.24±1.43)%,各组数据差异均有统计学意义(P<0.05)。0、50、100、200μmol/L白藜芦醇组S期细胞比例随着浓度增大而减小,分别为(34.89±0.66)%、(31.07±1.75)%,(28.66±1.71)%,(23.65±0.62)%,各组数据差异均有统计学意义(P<0.05)。免疫印迹法检测显示,白藜芦醇以时间依赖方式下调周期蛋白cyclin D1表达,上调p21cip1蛋白以及细胞凋亡相关蛋白剪切行Caspase-3和PRAP表达。结论白藜芦醇能明显抑制HCT-15细胞增殖,并诱导其细胞凋亡,可引起HCT-15细胞的S期阻滞。
Objective To investigate the effect of resveratrol on the cell proliferation and apoptosis of human colon cancer HCT-15cells.Methods Different concentrations of resveratrol(25,50,100,200umol/L)were used to treat human colon cancer HCT-15cells,non-treatment cells served as control group.MTT assay was used to detect the effect of resveratrol on the HCT-15cell proliferation.Morphological changes were observed under inverted phase contrast microscope.Flow cytometry was used to detect the cell cycle distribution and the percentage of apoptosis.Western blotting and polymerase chain reaction were used to observe the expression of major cell cycle proteins and genes expression.Results Resveratrol treatment significantly inhibited the growth of HCT-15cells in a time-and concentration-dependent manner.Cell morphology presented a shrinkage manner with the drug concentration increased.Flow cytometry showed that the percentage of G0/G1phase in human colon cancer HCT-15cells increased with the increasing of drug concentration for24h(0umol/L:42.81%±1.21%,50umol/L:49.25%±0.92%,100umol/L:54.67%±1.03%,200umol/L:64.24%±1.17%)and that it also increased at the concentration of50umol/L resveratrol with the treatment time(0h:32.41%±1.09%,24h:48.35%±1.03%,48h:64.24%±1.17%,72h:70.24%±1.43%),with statistically significant differences between control group and observation groups(all P<0.05).Resveratrol blocked the HCT-15cells in the S phase also in a dose-dependent manner(0umol/L:34.89%±0.66%,50umol/L:31.07%±1.75%,100umol/L:28.66%±1.71%,200umol/L:23.65%±0.62%),all with significant difference(P<0.05).The expression of cyclin D1was down-regulated by resveratrol in a time-dependent manner,but the expression of cleaved caspase-3,PARP and p21cip1proteins and genes were up-regulated.Conclusion Resveratrol can inhibit the proliferation of HCT-15cells and induce the cells to apoptosis by arresting the cells into S phase.
作者
胡旻
桑雅清
HU Min;SANG Yaqing(Division of Chinese Medicine, the Central Hospital of Lishui, Lishui(323000), China)
出处
《浙江中西医结合杂志》
2017年第12期1032-1034,共3页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
