7-二氟甲氧基-5,4′-二-正辛烷氧基金雀异黄素诱导肺癌A549细胞凋亡

Induction of apoptosis by 7-difluoromethoxyl-5,4'-di-n-octylgenistein in lung cancer A549 cells

目的研究7二氟甲氧基-5,4'二正辛烷氧基金雀异黄素(7-difluorom+hoxyl-5,4'-di-n-octylgenistein,DFOG)诱导人肺癌A549细胞凋亡作用。方法体外培养A549细胞。二氟甲基化或烷基化得到一系列金雀异黄素衍生物。MTT法测定细胞活力;碘化丙啶(PI)染色流式细胞术(FCM)检测细胞凋亡率;琼脂糖凝胶电泳观察DNA梯形条带;Western blot分析Bcl-2和Bax蛋白表达。结果 9个金雀异黄素衍生物较先导化合物GEN具有更强的抗肿瘤活性。其中DFOG对A549细胞活性显示最强的抑制作用,IC50为3.9μmol·L-1。DFOG(2.0、4.0和8.0μmol·L-1)作用48h的细胞凋亡率依次增高。8.0μmol·L-1DFOG处理A549细胞48h导致典型DNA梯形条带形成。8.0μmol·L1DFOG处理A549细胞6h、12h和24h,Bcl-2蛋白表达水平下降,而Bax蛋白表达上升。结论 DFOG诱导A549细胞凋亡作用与其增高Bax/Bcl-2比值有关。

Objective To investigate the effects of 7-difluoromethyl-5,4＇-di-n-octylgenistein （DFOG） on apoptosis of human lung caner A549 cells.Method A549 cells were cultured in vitro.A series of genistein derivatives were prepared by difluoromethylation and alkylation.The cell viability was measured by MTT assay.The apoptotic rate was detected using flow cytometry （FCM） after PI staining.DNA ladder bands were observed by DNA agarose gel electrophoresis.The expressions of Bcl-2 and Bax protein were analyzed by Western blot.Results Nine of genistein derivatives had more effective antitumor activity than genistein,among these derivatives,DFOG showed the strongest activity against A549 cells in vitro,and its IC50 was 3.9μmoL·L^-1.The apoptosis rates treated with 2.0,4.0 and 8.0μmol·L^-1 of DFOG for 48 h increased in turn.Treatment of A549 cells with 8.0μmo·L^-1 of DFOG for 48 h resulted in typical DNA ladder bands.Exposure of A549 cells to 8.0μmol·L^-1 DFOG at 6 h,12 h and 24 h,Bcl-2 protein level decreased,but Bax protein expression increased.Conclusion DFOG induces apoptosis of A549 cells is associated with increasing ratio of Bax to Bcl-2.