下调PAR基因表达对前列腺癌PC3细胞凋亡及凋亡相关蛋白Bcl-2/Bax的影响

Downregulation of PAR expression induces the apoptosis of human prostate cancer PC3 cells and increases the Bcl-2/Bax ratio

目的:本研究以RNA干扰技术下调PAR(prostate androgen regulated)基因表达,探讨其对前列腺癌PC3细胞增殖、凋亡及其相关蛋白Bcl-2/Bax表达水平的影响。方法:PC3细胞转染靶向PAR基因的siRNA,MTT法检测细胞增殖,用流式细胞术检测细胞凋亡,Western印迹检测Bcl-2和Bax蛋白表达水平。结果:PC3细胞PAR基因表达水平下调,此时处于G2-M期的细胞增加,为(29.95±3.25)%;细胞凋亡率亦明显增加,为(20.61±2.73)%,与对照组比较差异有显著性意义(P<0.01),同时Bcl-2蛋白表达下降,Bax表达水平升高,Bax/Bcl-2比值升高。结论:PAR基因表达下调可诱导细胞G2-M期阻滞和凋亡,其机制为抑制凋亡相关蛋白Bcl-2表达,同时促进Bax表达。

Objective： To investigate the effects of the downregulated expression of the prostate androgen regulated（PAR） gene on the cell cycle and apoptosis of PC3 cells as well as on the expression level of Bcl-2/Bax.Methods：After transfecting PC3 cells with small interfering RNA（siRNA） targeting PAR,we detected the inhibitory effect of PAR depletion on the proliferation of the PC3 cells by MTT assay,determined their apoptosis by flow cytometry,and measured the expression levels of Bcl-2 and Bax by Western blot.Results： The expression of PAR was suppressed by siRNA,the G2-M phase PC3 cells were increased to（29.95±3.25）%,and the apoptosis of the cells was enhanced to（20.61±2.73）%,with statistically significant difference from the control group（P0.01）.Western blot showed a decreased expression of Bcl-2,an increased expression of Bax,and an elevated ratio of Bax to Bcl-2.Conclusion： Downregulation of the PAR expression increases the Bax/Bcl-2 ratio and Bax expression,and thus induces the G2-M phase arrest and apoptosis of PC3 cells.