摘要
目的:探讨脂质体介导的短发夹RNA(shRNA)对前列腺癌细胞中PIM-1基因表达的影响。方法:构建3种针对PIM-1mRNA不同靶点的shRNA重组质粒表达载体,并转染前列腺癌PC-3细胞。分别采用逆转录-聚合酶链反应(RT-PCR)及蛋白印迹法(Westernblot)检测转染后细胞中PIM-1mRNA及蛋白的表达情况。结果:转染48h后,3种PIM-1基因靶向性shRNA表达质粒均可抑制PC-3细胞中PIM-1mRNA和蛋白的表达,其中靶向PIM-1mRNA第707-725位和1080-1098位序列的重组质粒干扰效果更显著。结论:本研究所构建的PIM-1shRNA表达质粒可在体外高效特异地抑制前列腺癌细胞中PIM-1的表达,为下一步研究奠定了基础。
Objective： To investigate the interference effect of lipofectamine-mediated short hairpin RNA （shRNA） on the expression of PIM-1 gene in prostate cancer cell lines. Methods： Three kinds of recombinant plasmid expression vectors （p^PIM1-shRNA-1,-2,-3） were constructed, which can produce shRNA targeting different sequence of PIM-1 mRNA. The recombinant plasmid vectors were transfected into PC-3 cell lines, respectively. RT-PCR and Western blot assay were used to detect the expression of PIM-1 mRNA and protein after transfection. Results： The expression of mRNA and protein of PIM-1 in PC-3 were inhibited significantly at 48h post-transfection. Furthermore, p^PIM1-shRNA-2 and p^PIM1-shRNA-3 resulted in a stronger silencing effect than that of p^PIM1-shRNA-1. Conclusion： The recombinant plasmid expression vector which carrying PIM-1 shRNA may remarkably inhibit the expression of PIM-1 mRNA and protein in PC-3 cell lines. The experiment provided the basis for further investigation of the role of PIM-1.
出处
《天津医药》
CAS
北大核心
2009年第7期529-531,I0001,共4页
Tianjin Medical Journal
基金
天津市科委科技支撑项目(项目编号:07ZCGYSF01000)
