Bax对人卵巢癌细胞的凋亡诱导作用及其机制研究

发布时间：2018-05-23 06:36

本文选题：Bax + 线粒体　；参考：《四川大学学报(医学版)》2015年05期

【摘要】：目的观察Bax对人卵巢癌A2780细胞的凋亡诱导作用,并探讨其作用机制。方法构建重组质粒pcDNA-Bax,将重组质粒pcDNA-Bax转染A2780细胞,采用Hoechst 33258染色观察细胞凋亡、MTT法检测细胞活力,采用Western blot法检测Bax基因的过表达、线粒体细胞色素C的释放及Caspase-9和Caspase-3的激活。结果经酶切和测序鉴定,重组质粒pcDNA-Bax构建成功。转染质粒pcDNA-Bax能明显诱导A2780细胞凋亡,Hoechst染色显示凋亡的细胞出现细胞核形态学改变;MTT结果显示转染质粒pcDNA3.1-Bax后,细胞活力明显下降;Western blot结果表明转染质粒pcDNA-Bax后的A2780细胞线粒体释放细胞色素C,激活了Caspase-9和Caspase-3。结论 Bax能促进卵巢癌细胞线粒体细胞色素C释放,从而诱导细胞凋亡。
[Abstract]:Objective to investigate the apoptosis-inducing effect of Bax on human ovarian cancer cell line A 2780 and its mechanism. Methods the recombinant plasmid pcDNA-Baxwas constructed. The recombinant plasmid pcDNA-Bax was transfected into A2780 cells. The viability of A2780 cells was detected by Hoechst 33258 staining. The overexpression of Bax gene, the release of mitochondrial cytochrome C and the activation of Caspase-9 and Caspase-3 were detected by Western blot assay. Results the recombinant plasmid pcDNA-Bax was successfully constructed by enzyme digestion and sequencing. Transfection of plasmid pcDNA-Bax could induce apoptosis of A2780 cells by Hoechst staining. The results of Western blot showed that the mitochondria of A2780 cells transfected with plasmid pcDNA-Bax released cytochrome C and activated Caspase-9 and Caspase-3. Conclusion Bax can promote the release of cytochrome C from mitochondria of ovarian cancer cells and induce apoptosis.
【作者单位】：成都医学院学报编辑部;湖北医药学院微生物学教研室;成都医学院临床医学实验教学中心;成都医学院病原生物学教研室;
【基金】：四川省教育厅面上项目(No.11ZB170) 四川省卫生厅科研课题(No.120489) 成都医学院校基金(No.CYZ10-004,No.CYZ11-0059)资助
【分类号】：R737.31

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