毛蕊异黄酮-7-O-β-D-葡萄糖苷对人宫颈癌HeLa细胞凋亡及Bcl-2/Bax表达的影响

Effects of calycosin-7-O-β-D-glucoside on cell apoptosis in cervical cancer HeLa cells and expression of Bcl-2/Bax

目的探讨毛蕊异黄酮-7-O-β-D-葡萄糖苷（CG）对宫颈癌细胞系HeLa细胞凋亡及Bcl-2／Bax表达的影响。方法体外培养HeLa细胞，分为对照组和实验组，实验组用不同质量浓度的CG分别处理24、48h，采用MTT法测定CG对HeLa细胞增殖的影响；Hoechst 33258染色观察细胞凋亡情况；采用流式细胞仪检测细胞凋亡和周期;Western blotting法检测半胱氨酸天冬氨酸蛋白酶（Caspase-3）、Bcl-2／Bax蛋白表达情况。结果CG（2．5-100μg／mL）能抑制HeLa细胞的增殖，呈剂量和时间依赖性；质量浓度为20、40、80μg／mL的CG作用HeLa细胞48h后，细胞凋亡率逐渐上升，分别为10．40％、25．50％、39．40％，明显高于对照组（1．80％）；实验组HeLa细胞Caspase-3的活性形式cleaved Caspase表达量显著增加；促凋亡蛋白Bax表达量升高，而抗凋亡蛋白Bcl-2表达量降低，呈剂量依赖性。结论CG能抑制HeLa细胞的增殖并诱导其凋亡，其作用机制可能与增加细胞Caspase-3活性、下调Bcl-2蛋白的表达及上调Bax蛋白有关。

Objective To explore the effect of calycosin-7-O-β-D-glucoside （CG） on apoptosis in cervical cancer HeLa cells and expression of Bcl-2/Bax. Methods HeLa cells were cultured and divided into two groups, including control group and experimental group. Cell viabilities were determined by the MTT method; Apoptosis and cell cycle were analyzed by flow cytometry; The changes of protein expression of cleaved Caspase-3, Bcl-2, and Bax were detected by Western blotting. Results From the data of MTT, the cell proliferation of human cervical cancer HeLa cells was inhibited by CG （2.5-100 μg/mL） in a dose- and time-dependent manner. Flow cytometry assays showed that EGCG significantly induced the apoptosis in HeLa cells. The apoptosis rate of the experimental group were increased gradually in 48 h after treated with CG （20, 40, and 80 μg/mL）, they were 10.40%, 25.50%, and 39.40%, respectively, significantly higher than those in the control group. The data of Western blotting showed that CG down-regulated Bcl-2 and up-regulated cleaved Caspase-3 and Bax in a dose-dependent manner. Conclusion CG could inhibit the the proliferation of HeLa cells and promote apoptosis, and the anticancer effect of CG may be associated with the down-regulation of Bcl-2 expression and up-regulation of Bax expression, as well as the increase of relative activity of Caspase-3. CG may be a promising antitumor agent for cancer treatment.