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目的:探讨米非司酮(RU486)对人前列腺癌PC-3和LNCap细胞增殖及相关基因CyclinE、CDK2、p21和p27表达的影响。方法:体外培养前列腺癌PC-3和LNCap细胞,用RU486处理PC-3和LNCap细胞,CCK-8检测对细胞增殖的影响;流式细胞仪检测对细胞周期的影响;RT-PCR法检测细胞增殖相关基因CyclinE、CDK2、p21和p27mRNA的表达,蛋白质印迹法检测细胞增殖相关蛋白CDK2、pCDK2的表达。结果:RU486对PC-3和LNCap细胞体外增殖均有明显的抑制作用,且呈时间-剂量依赖性;25μmol/LRU486处理PC-3和LNCap细胞24h后,2种细胞均被阻滞在G1/S期;不同浓度的RU486处理PC-3和LNCap细胞24h后,随着RU486浓度的增加,Cyclin E mRNA表达下降,p21和p27mRNA表达增加,CDK2 mRNA表达无明显变化,但pCDK2表达呈下降趋势。结论:RU486能使前列腺癌LNCap和PC-3细胞周期阻滞在G1/S期,并能通过上调p21和p27mRNA的表达,下调Cyclin E mRNA及磷酸化的CDK2蛋白而抑制细胞增殖。
Objective: To investigate the effects of mifepristone (RU486) on the proliferation of PC-3 and LNCap cells and the expression of CyclinE, CDK2, p21 and p27 in human prostate cancer cells. Methods: PC-3 and LNCap cells were cultured in vitro. The effects of RU486 on PC-3 and LNCap cells and the effect of CCK-8 on cell proliferation were detected by flow cytometry. The cell cycle was detected by RT-PCR The expression of CyclinE, CDK2, p21 and p27 mRNA were detected by Western blotting. The expressions of CDK2 and pCDK2 were detected by Western blot. Results: RU486 inhibited the proliferation of PC-3 and LNCap cells in vitro in a time-and dose-dependent manner. After treated with 25μmol / L LR488 for 24 h, PC-3 and LNCap cells were arrested in G1 / S. After treatment with different concentrations of RU486 for 24 h, the expression of Cyclin E mRNA decreased and the expression of p21 and p27 mRNA increased but the expression of pCDK2 decreased with the increase of RU486 concentration. Conclusion: RU486 can block the cell cycle of LNCap and PC-3 cells in G1 / S phase of prostate cancer and inhibit cell proliferation by up-regulating the expression of p21 and p27 mRNA, down-regulating Cyclin E mRNA and phosphorylating CDK2.