RNA干扰PTEN基因对HCT-8细胞增殖能力的影响  

Influence of PTEN on proliferation of colon cancer cell HCT-8 by siRNA

作　　者：吕强[1] 唐亮[2] 李建华[1] 宫鹏涛[1] 徐晓芳[3] 田甜[1] 李泽中[3] 雒伟伟[3] 邢沈阳[3] 张西臣[1] 高久春[4] 

机构地区：[1]吉林大学畜牧兽医学院预防兽医学系,吉林长春130062 [2]吉林农业大学动物科技学院预防兽医学系,吉林长春130118 [3]吉林大学畜牧兽医学院动物遗传育种与繁殖系,吉林长春130062 [4]吉林大学白求恩医学院生物学教研室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2011年第2期231-235,394,共5页Journal of Jilin University：Medicine Edition

基　　金：国家重大基础研究计划项目资助课题(2006CB910505)

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。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.

关 键 词：PTEN 人结肠癌HCT-8细胞 小于扰RNA 

分 类 号：Q752[生物学—分子生物学] R735.35[医药卫生—肿瘤]