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

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24h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.

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

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<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<0.05 or P<0.01). EGB could enhance MCF-7 cells Caspases-3 protein expression (P<0.05 or P<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.