金雀异黄素对胃癌细胞增殖和细胞周期的影响

Effect of genistein on proliferation and cell cycles of gastric cancer cells

目的 酪氨酸蛋白激酶抑制剂——金雀异黄素对人胃癌细胞MGC-803的抑制作用,及其对细胞周期素（Cyclin） B1蛋白表达和细胞周期的影响.方法 体外培养MGC-803细胞,经170、200、220μmol/L的金雀异黄素分组处理后采用噻唑蓝（MTT）法判断金雀异黄素对MGC-803的抑制作用;流式细胞仪检测细胞周期的变化;采用Western blot法检测Cyclin B1和周期蛋白依赖性蛋白激酶1（CDK1）蛋白的表达.结果 MTT法检测显示随着金雀异黄素浓度增高,作用时间24-72 h的延长,MGC-803细胞增殖抑制率升高,呈时间和剂量依赖性（P＜0.01）.流式细胞术结果显示24、48、72 h金雀异黄素组与对照组比较均出现S期阻滞[（24.37±3.25）％、（27.11±2.82）％、（28.20±4.17）％比（12.35±2.62）％,P＜0.01]和G2/M期阻滞[（23.11±3.31）％、（27.41±2.70）％、（35.61±2.67）％比（9.65±3.62）％,P＜0.01],其中以72 h组的G2/M期细胞比例的增加最为显著.Western blot结果显示,随着金雀异黄素浓度增高（170、200、220μmmol/L）,处理时间的延长（24、48、72 h）,Cyclin B1和CDK1蛋白表达均逐渐减少,与对照组比较差异有统计学意义（P＜0.01）.结论 金雀异黄素可以有效地降低Cyclin B1和CDK1蛋白的表达,同时对MGC-803细胞的细胞周期也会产生影响,金雀异黄素对MGC-803细胞的增殖起抑制作用.

Objective To investigate the inhibitory effect of histidine protein kinase inhibitor genistein on human gastric cancer cell line MGC-803,and its effect on the expression of Cyclin B1 and cell cycle.Methods MGG803 cells were cultured.Methyl thiazol tetrazolium （MTT） assay was used to examine the inhibitory effect of genistein in MGC803 cells.Three concentrations of genistein were used in MTT assay：170,200,and 220 μ mol/L.Western blotting was used to detect the expression of Cyclin B1 and CDK1.Results With the increase of genistein concent ration and prolongation of time,from 24 to 72 h,MTT showed the inhibitory rate of proliferation of MGC-803 cells increased in a dose-and time-dependent manner （P 〈0.01）.Flow cytometry indicated that as compared with control group,the S percentage in 24,48 and 72 h genistein groups was （24.37 ±3.25）%,（27.11 ±2.82）% and （28.20 ±4.17）%respectively vs.（12.35 ±2.62）% （P〈0.01）,and G2/M percentage was （23.11±3.31）%,（27.41 ±2.70）% and （35.61 ±2.67）% respectively vs.（9.65 ±3.62）% （P〈0.01）.When MGC-803 cells were treated with genistein for 72 h,the cell rate at G2/M were the most significantly increased.Western blot showed that,with the increase genistein concent ration （170,200 and 220 μmmol/L） and prolongation of time （24,48 and 72 h）,the expression of protein Cyclin B1 and CDK1 gradually decreased.There are significant differences by compared with the control group （P 〈 0.01）.Conclusion Genistein can decrease the expression of Cyclin B1 and CDK1 in MGC-803 cells,and also could inhibit cell cycle and proliferation of MGC803 cells.