目的 探讨红三叶异黄酮对人宫颈癌HeLa细胞的抑制作用及其作用机制。方法 以MTT法检测红三叶异黄酮20、40、80 μg/mL分别在24、48、72 h对人宫颈癌HeLa细胞增殖作用的影响，并在倒置显微镜下观察HeLa细胞形态学改变。流式细胞仪检测红三叶异黄酮20、40、80 μg/mL对HeLa细胞凋亡的影响。实时荧光定量PCR仪检测红三叶异黄酮20、40、80 μg/mL对HeLa细胞凋亡中调节基因Bax、Bcl-2表达水平的影响。结果 红三叶异黄酮对人宫颈癌HeLa细胞的增殖抑制作用，表现为时间、浓度的相关性。红三叶异黄酮对HeLa细胞的形态学改变，表现为随药物浓度增加细胞固缩变小，并出现凋亡小体。流式细胞仪检测分析显示红三叶异黄酮能引起HeLa细胞凋亡，并随药物浓度的增加而明显，当红三叶异黄酮达到一定有效浓度时，诱导其细胞凋亡。通过调节Bax mRNA与Bcl-2 mRNA的表达，促进抑制HeLa细胞增殖，能使Bax表达上升、Bcl-2表达下降，且呈剂量相关性。结论 红三叶异黄酮对人宫颈癌HeLa细胞的增殖具有抑制作用，并呈一定的剂量和时间相关性，提示通过调节Bax mRNA与Bcl-2 mRNA的表达，促进抑制HeLa细胞增殖。

Objective To investigate the effects and mechanism of red clover isoflavones on inhibition of human cervical cancer HeLa cells. Methods The MTT assay was used to detect the effects of red clover isoflavones with concentrations of 20, 40, and 80 μg/mL, respectively, on the proliferation of human cervical cancer HeLa cells at 24, 48, and 72 h, respectively. The morphological changes of HeLa cells were observed under inverted microscope. Flow cytometry was used to detect the apoptotic rate of red clover isoflavones with concentrations of 20, 40, and 80 μg/mL, respectively on HeLa cells. Real-time PCR instrument was used to detect red clover isoflavones with concentrations of 20, 40, and 80 μg/mL on HeLa cells apoptosis modulating genes Bax and Bcl-2 expression levels. Results Red clover isoflavones inhibited the proliferation of human cervical cancer HeLa cells in a time and concentration dependent manner. The morphological changes of HeLa cells in red clover isoflavones showed that with the increase of drug concentration, cell shrinkage decreased and apoptotic globules appeared. Flow cytometry analysis showed that red clover isoflavones could induce apoptosis of HeLa cells, which was obvious with the increase of drug concentration. When it reached a certain effective concentration, it could induce apoptosis of HeLa cells. By regulating the expression of mRNA Bax and mRNA Bcl-2 to promote the inhibition of HeLa cell proliferation, Bax expression was increased, while Bcl-2 expression was decreased, and was dose dependent. Conclusion Red clover isoflavones has the inhibition of human cervical cancer HeLa cells in a dose and time dependent manner, indicating that red clover isoflavones promote the inhibition of HeLa cell proliferation by regulating the expression of mRNA Bax and mRNA Bcl-2.

天津市"131"创新型人才培养工程（2016）