摘要
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 impact of RU486 on proliferation of the prostate cancer cells and on the expression of cell cycle related genes like Cyclin E,p21,p27 and CDK2,in PC-3 and LNCap cell lines. METHODS：LNCap and PC-3 prostate cancer cell lines were cultured at different concentrations of RU486 in vitro. The effects of RU486 on the proliferation of cells were analyzed by CCK-8 assay. Cell cycle analysis was measured by FACS. The mRNA expression levels of Cyclin E,p21,p27 and CDK2 were examined by RT-PCR. The protein expression levels of CDK2 and pCDK2 were measured by Western blotting. RESULTS：The treatment of both cells with RU486 inhibited cell proliferation in a time-and dose-dependent manner by CCK-8 assay. Analysis of the cell cycle with FACS demonstrated that the treatment of both cells with 25 μmol/L RU486 24 h induced cell-growth arrest at G1/S phase. The increasing concentration of RU486 caused a marked increase in p21 and p27 mRNA levels and a significant decrease in Cyclin E mRNA level in both cell lines following 24 h exposure to different concentration of RU486. Moreover,it did not alter the expression of CDK2 mRNA,but reduced the expression of pCDK2 protein in both cells. CONCLUSION：RU486 can induce G1/S arrest and has a potent anti-proliferative effect on prostate cancer cells through the p21 and p27 mRNA up-regulation,and Cyclin E mRNA and pCDK2 protein down-regulation.
出处
《中华肿瘤防治杂志》
CAS
2010年第17期1328-1331,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
黑龙江省教育厅科学技术研究面上项目(1153108)
哈尔滨市科技创新人才研究专项资金项目(2007RFXXS039)
关键词
米非司酮
前列腺肿瘤
细胞增殖
基因表达
mifepristone
prostatic neoplasms
cell proliferation
gene expression
