金雀异黄酮对三阴乳腺癌MDA-MB-231细胞凋亡及EGFR/PI3K/Akt通路的影响

摘要

Aim To investigate the effect of genistein on apoptosis in triple-negative breast cancer MDA-MB-231 cells and the underlying mechanisms.Methods MTT assay was used to detect the inhibition rate on breast cancer MDA-MB-231 cells of genistein.Hoechst 33258 staining was applied to determine the effect of genistein on morphology of MDA-MB-231 cells.qRT-PCR was employed to detect the mRNA expression of EGFR in MDA-MB-231 cells.Western blot was utilized to determine the expression of Bcl-2, Bax, caspase-3, EGFR, Akt, and p-Akt.The expressions of Akt and p-Akt proteins in breast cancer MDA-MB-231 cells were detected after treated with Akt activator insulin, genistein and in combination with insulin.Results Genistein inhibited the viability of breast cancer MDA-MB-231 cells in a time-dependent manner.The results of Hoechst 33258 staining showed a typical apoptotic morphological changes of MDA-MB-231 cells after treatment of genistein for 36 h.qRT-PCR showed that the mRNA expression of EGFR in MDA-MB-231 cells decreased after treated with genistein for 36 h.The expression levels of Bcl-2, EGFR, Akt, p-Akt, ERK, p-ERK were significantly down-regulated(P<0.01) compared with control.While, the expression of Bax, caspase-3 was significantly up-regulated (P<0.01).It was observed that p-Akt was significantly activated after the treatment of Akt activator insulin (P<0.01), however, significantly down-regulated (P<0.01) when treated with genistein.Conclusion Genistein could inhibit the growth of triple-negative breast cancer MDA-MB-231 cells and induce apoptosis, which probably involves regulating EGFR/PI3K/Akt signaling pathway.%目的 研究金雀异黄酮(genistein)诱导三阴性乳腺癌MDA-MB-231细胞凋亡及其机制. 方法 MTT法观察金雀异黄酮对乳腺癌MDA-MB-231细胞增殖的抑制作用;Hoechst 33258染色观察金雀异黄酮对MDA-MB-231细胞核凋亡形态学的影响;qRT-PCR法观察金雀异黄酮干预MDA-MB-231细胞36 h后,EGFR mRNA表达水平的变化;金雀异黄酮干预MDA-MB-231细胞36 h后,Western blot检测凋亡相关蛋白Bcl-2、Bax、caspase-3,EGFR、Akt、p-Akt蛋白的变化;Akt激活剂胰岛素(insulin)、金雀异黄酮单独及联合胰岛素干预乳腺癌MDA-MB-231细胞后,Western blot检测Akt和p-Akt蛋白表达量的变化. 结果 MTT结果显示,金雀异黄酮呈时间浓度依赖性抑制乳腺癌MDA-MB-231细胞增殖;Hoechst 33258染色结果显示,金雀异黄酮干预乳腺癌MDA-MB-231细胞36 h后细胞核呈现典型凋亡形态学改变;qRT-PCR结果显示,经金雀异黄酮干预MDA-MB-231细胞36 h后,EGFR的mRNA表达水平明显下降(P<0.01);Western blot结果显示金雀异黄酮干预乳腺癌MDA-MB-231细胞36 h后,与对照组对比,Bcl-2、EGFR、Akt、p-Akt蛋白表达水平明显下调(P<0.01),Bax、caspase-3蛋白表达水平明显上调(P<0.01),Akt激活剂胰岛素可以明显激活p-Akt(P<0.01),金雀异黄酮可以明显下调被激活的p-Akt(P<0.01). 结论 金雀异黄酮能抑制三阴乳腺癌MDA-MB-231细胞生长并诱导其凋亡,其机制可能与抑制EGFR/PI3K/Akt信号通路有关.