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目的研究金雀异黄素(Genistein,Gen)对人乳腺癌细胞株MCF-7凋亡的影响及凋亡相关基因表达的调控。方法选用人乳腺癌细胞株MCF-7体外培养,MTT法检测Gen对乳腺癌生长活力的影响,流式细胞仪检测细胞周期及凋亡率,Giemsa染色观察细胞形态学改变,免疫组化检测凋亡相关基因Bax和癌基因erbB-2蛋白表达水平的改变。结果金雀异黄素对MCF-7细胞体外生长具有明显的抑制作用。Giemsa染色显示,金雀异黄素处理的MCF-7细胞呈明显的凋亡形态改变,细胞固缩、发泡,染色质凝集呈块状,并沿核周分布。流式细胞仪检测在G1峰前可见凋亡峰,且随用药时间的延长,凋亡比率递增。蛋白水平检测表明金雀异黄素促进MCF-7细胞Bax表达升高,抑制erbB-2蛋白的表达。结论金雀异黄素通过诱导细胞凋亡或阻滞细胞于G2～M期,从而抑制乳腺癌细胞的体外增殖,并主要是通过调节Bax和erbB-2蛋白的表达实现的。
Objective To investigate the effects of Genistein on the apoptosis of human breast cancer cell line MCF-7 and the regulation of apoptosis related gene expression. Methods The human breast cancer cell line MCF-7 was cultured in vitro. The effect of Gen on the growth of breast cancer cells was detected by MTT assay. The cell cycle and apoptosis rate were detected by flow cytometry. The morphological changes were observed by Giemsa staining. Changes in expression of Bax and oncogene erbB-2. Results Genistein had a significant inhibitory effect on the growth of MCF-7 cells in vitro. Giemsa staining showed that genistein-treated MCF-7 cells showed obvious apoptotic morphological changes, cell shrinkage, foaming, chromatin agglutination in blocks, and along the perinuclear weeks. The apoptotic peak was observed before G1 peak by flow cytometry, and the apoptosis rate increased with the prolongation of drug use time. Protein levels showed that genistein enhanced the expression of Bax in MCF-7 cells and inhibited the expression of erbB-2 protein. Conclusion Genistein can inhibit the proliferation of breast cancer cells in vitro by inducing apoptosis or arresting cells at G2 ~ M phase, and mainly through the regulation of the expression of Bax and erbB-2 proteins.