摘要目的 探讨中介体复合物亚基19(Med19)对人前列腺癌PC3细胞侵袭迁移能力的影响及其在上皮-间充质(EMT)转化过程中的机制.方法 采用针对Med19的siRNA慢病毒载体感染PC3细胞,应用实时荧光定量PCR(qRT-PCR)和蛋白质印迹方法(Western印迹)检测Med19-siRNA感染组(siRNA)与空载组(scRNA)Medl9基因表达情况;采用Boyden小室侵袭实验、划痕实验检测感染后PC3细胞侵袭、迁移能力变化;采用qRT-PCR检测感染后PC3细胞上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、增强子结合蛋白-2(ZEB2)、转录因子Snail-1和Snail-2 mRNA的表达.结果 Medl9-siRNA慢病毒感染PC3细胞后,实验组Medl9 mRNA及蛋白表达水平(16.9±3.4)％比空载组近100％明显降低,差异有统计学意义(P<0.01),Boyden小室侵袭实验和划痕实验显示感染组细胞侵袭、迁移能力明显减弱(P<0.01),感染组N-Cadherin、Vimentin、ZEB2、Snail-1和Snail-2表达降低,E-Cadherin表达升高,差异有统计学意义(P<0.01).结论 沉默Med 19基因可通过抑制EMT中的相关基因,降低人前列腺癌PC3细胞的侵袭转移能力.

abstractsObjective To studythe effect of lentivirus-mediated inhibition of Med19 on cell migration andinvasion in the PC3 cells, and explore the mechanism of epithelial-mesenchymal transition transformation. Methods The lentivirus vectors containing small interferingRNA(siRNA) targeting Medl9 gene were constructed and transfected to PC3 cells.Quantitative real-time PCR(qRT-PCR) and Western blot analysis were used to detect the Medl9 expression in the siRNA group(PC3-Med 19-siRNA cells)and the NC group(PC3-Med 19-sc cells) at 72h after the transfection. The cell mobility, migration and invasion ability of PC3 cells were respectively measured by Boyden migration and woun-healing assay. The expression of E-Cadherin, N-Cadherin, Vimentin, ZEB2, Snail-1, and Snail-2 mRNA were detected by using qRT-PCR. Results The expression of Medl9 mRNA in PC3-Med 19-siRNA cells was lower than that in PC3-Med 19-scRNA cells(P<0.01). The number of migrated cells and invaded cells were significantly decreased in PC3-Med 19-siRNA cells(P<0.01). The expression of N-Cadherin, Vimentin, ZEB2, Snail-1, and Snail-2 mRNA were remarkablylower and E-Cadherin was higher in PC3-Med 19-siRNA cells. Conclusion Med 19 inhibitioncouldreduce migration abilityof prostate cancer PC3 cells by epithelial-mesenchymal transition.