银杏叶提取物对乳腺癌MCF-7细胞增殖、凋亡及Caspase-3表达的影响

Effects of Ginkgo biloba Extract on Proliferation,Apoptosis and Caspase-3 Expression in Human Breast Cancer MCF-7 Cells

目的:探讨银杏叶提取物(EGB)对乳腺癌MCF-7细胞增殖、凋亡及半胱氨酸蛋白酶(Caspases)-3表达的影响。方法:将质量浓度为0,10,20,40,80,160 mg·L-1的EGB作用于体外培养的人乳腺癌MCF-7细胞上,培养24 h或48 h,然后应用四甲基偶氮唑蓝染色法(MTT法)检测细胞增殖;Annexin V/PI双染色流式细胞仪检测细胞凋亡和细胞周期;酶联免疫吸附法(ELISA)检测Caspases-3蛋白表达。结果:EGB对MCF-7细胞的体外增殖具有抑制作用,量效关系显著,与对照组比较有统计学差异(P<0.01),半数抑制浓度(IC50)为83.65 mg·L-1。经流式细胞仪检测表明,EGB能使MCF-7细胞G0-G1期逐渐增加,G2-M期和S期逐渐减少,并且随着质量浓度的增加,MCF-7细胞凋亡率明显增加(P<0.05或P<0.01)。EGB能增强MCF-7细胞Caspases-3蛋白的表达(P<0.05或P<0.01),并呈浓度依赖性。结论:EGB能有效抑制人乳腺癌MCF-7细胞Caspases-3蛋白表达,诱导MCF-7细胞凋亡,抑制肿瘤细胞增殖。

Objective：To investigate the effects of Ginkgo biloba extract（EGB） on proliferation,apoptosis and Caspase-3 expression in human breast cancer MCF-7 cells.Method：The human breast cancer MCF-7 cells cultivated in vitro were treated with 0,10,20,40,80,160 mg·L-1 of EGB,cultured for 24 h or 48 h.The 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl-tetrazolium bromide（MTT） assay was used to measure the cell proliferative effect.The annexin V/PI double staining analysis by flow cytometry was used to evaluate apoptotic rate and the cell cycle.The expression of Caspases-3 protein was detected by enzyme-linked immunosorbent assay（ELISA）.Result：EGB could inhibit the proliferation of MCF-7 cells in vitro,and concentration-effect relationship was significant（P ＆lt; 0.01）.50% inhibition concentration（IC50） of MCF-7 was 83.65 mg·L-1.The flow cytometry test indicated that EGB could make MCF-7 cells G0-G1 phase gradually increasing,G2-M period and S phase reduce gradually and with the increase of the concentration,MCF-7 cell apoptosis rate increased significantly（P ＆lt; 0.05 or P ＆lt; 0.01）.EGB could enhance MCF-7 cells Caspases-3 protein expression（P ＆lt; 0.05 or P ＆lt; 0.01） and with the concentration dependent.Conclusion：EGB could effectively restrain human breast cancer MCF-7 cells Caspases-3 protein expression,induction MCF-7 cell apoptosis,inhibiting tumor cell proliferation.