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[摘要]
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、48 h, 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(2.5~100 μg/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、80 μg/mL的CG作用HeLa细胞48 h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。
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[Abstract]
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.
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