【摘要】 目的:观察小干扰RNA(siRNA)对结肠癌相关基因PTEN表达的抑制作用及其对结肠癌细胞增殖能力的影响。方法:构建携带有针对PTEN基因序列的siRNA真核绿色荧光表达载体重组质粒pGPU6/GFP/Neo-PTEN-1、pGPU6/GFP/Neo-PTEN-2、pGPU6/GFP/Neo-PTEN-3和pGPU6/GFP/Neo-PTEN-4,脂质体法转染入结肠癌HCT-8细胞(分别为p1、p2、p3和p4组),同时设转染阴性对照质粒组(p5组)和亲本细胞组(p6组),实时PCR方法检测PTEN mRNA表达,MTT法分析细胞增殖活性变化。结果:通过荧光显微镜观察计数,细胞转染率为58%。实时PCR方法检测结果显示,siRNA重组质粒pGPU6/GFP/Neo-PTEN-1、pGPU6/GFP/Neo-PTEN-2、pGPU6/GFP/Neo-PTEN-3和pGPU6/GFP/Neo-PTEN-4的抑制率分别为48.3%、76.5%、29.4%和61.2%,与p6组比较差异均有统计学意义(P<0.05)。MTT结果表明,细胞增殖抑制能力明显增强。结论:PTEN基因在肿瘤的发生发展中可能起到一定的作用。更多还原

【Abstract】 Objective To observe the inhibitory effect of small interfering RNA(siRNA) on the expression of colorectal cancer related gene PTEN and the influence in the proliferation of colon cancer cells in vitro.Methods The recombinant plasmids which contained the siRNA of the PTEN gene in the green fluorescence eukaryotic vectors(pGPU6/GFP/Neo-PTEN-1,pGPU6/GFP/Neo-PTEN-2,pGPU6/GFP/Neo-PTEN-3 and pGPU6/GFP/Neo-PTEN-4) were constructed and then they were transfected into colon cancer cells by using Lipofectamine 2 000.The negative control(p5) group and parent cells(p6)group were set up at the same time.The expression of PTEN gene mRNA was detected by real-time PCR.The proliferation of colon cancer cells was detected by MTT.Results Counted by fluorescence microscopy,the transfection rate of cells was 58%.The inhibitory rates of PTEN mRNA in the pGPU6/GFP/ Neo-PTEN-1,pGPU6/GFP/ Neo-PTEN-2,pGPU6/GFP/Neo-PTEN-3 and pGPU6/GFP/Neo-PTEN-4 were 48.3%,76.5%,29.4%,and 61.2%,there were significant differences compared with p6 group(P<0.05).The proliferation abilities of colon cells were significantly inhibited.Conclusion PTEN gene may play a role in the development of colorectal cancer.更多还原