20(s)-原人参二醇对体外培养的人前列腺癌PC3细胞凋亡的诱导作用及其机制

Inductive effect of 20(s)-proto-panaxdiol on apoptosis of human prostate cancer PC3 cells cultivated in vitro and its mechanism

目的:观察20(s)-原人参二醇(PPD)对体外培养的人前列腺癌(PCa)PC3细胞凋亡的作用,阐明其抗肿瘤作用机制。方法:将体外培养的PC3细胞分为对照组及20、30和40μmol.L-1 PPD组。采用PPD处理细胞48h后,利用光学显微镜观察细胞形态学改变,MTT法检测细胞增殖抑制情况,采用吖啶橙/溴化乙锭(AO/EB)染色法检测细胞凋亡情况,并通过Western blotting方法检测Bcl-2、Bax和γH2Ax蛋白的表达情况。结果:不同剂量PPD处理后,PC3细胞变圆回缩并出现碎片,AO/EB染色呈现不同程度的黄绿色;不同剂量PPD作用后,各组细胞增殖抑制率均显著升高,与对照组比较差异有统计学意义(P<0.05);不同剂量PPD作用PC3细胞48h后,Bcl-2蛋白表达明显下调(P<0.05),Bax蛋白表达变化不明显,γH2Ax蛋白表达上调(P<0.05)。结论:PPD可诱导PC3细胞凋亡,其作用机制可能与Bcl-2、Bax信号通路以及DNA断裂基因γH2Ax蛋白表达上调有关。

Objective To study the effect of 20（s）-proto-panaxdiol（PPD） on apoptosis of human prostate cancer PC3 cells cultivated in vitro,and to clarify the mechanism of its anticancer action.Methods The PC3 cells cultivated in vitro were divided into control,and 20,30,40 μmol·L-1 PPD groups.The morphological changes of PC3 cells were observed under optical microscope after treated with PPD for 48 h.The proliferation inhibition of PC3 cells was detected by MTT method.The apoptosis of PC3 cells was detected by AO/EB staining and the expressions of Bax,Bcl-2 and γH2Ax proteins were analyzed by Western blotting.Results After treated with different doses of PPD,the PC3 cells were round,recovery,and broken.AO/EB staining found that the cells presented yellow-green.The inhibitory rates of proliferation of PC3 cells in various groups were increased compared with control group（P〈0.05）.The expression of Bcl-2 protein was down-regulated（P〈0.05）,the expression of Bax protein didn’t change significantly,and the expression of γH2Ax protein was up-regulated（P〈0.05）.Conclusion PPD may induce the apoptosis of PC3 cells,and its mechanism may be related to the down-regulation of Bcl-2 protein and the up-regulation of γH2Ax protein expression.