siRNA干扰人端粒酶逆转录酶对胃癌细胞体外增殖转移的抑制作用  

Inhibitory Effect on in vitro Proliferation and Migration of Gastric Cancer Cells of Silencing Human Telomerase Reverse Transcriptase by SiRNA

作　　者：徐航[1] 邓家齐[2] 吕沐瀚[3] 

机构地区：[1]泸州医学院附属中医医院心脑电图室,四川泸州646000 [2]泸州医学院附医学英语教研室,四川泸州646000 [3]泸州医学院附属医院消化内科,四川泸州646000

出　　处：《华南国防医学杂志》2013年第10期699-702,714,共5页Military Medical Journal of South China

基　　金：2011年泸州医学院科研基金(12267);2012年泸州医学院附院课题(12048)

摘　　要：目的利用RNA干扰(RNA interference,RNAi)技术构建人端粒酶逆转录酶(human telomerase reverse transcriptase,hTERT)小干扰RNA(small interfering RNA,siRNA)化学合成片段,观察hTERT基因干扰后,胃癌SGC-7901细胞增殖及迁移能力的变化。方法在高表达hTERT的胃癌细胞株SGC-7901中转染hTERT siRNA,经荧光显微镜验证转染成功,实时聚合酶联反应(real-time polymerase chain reaction,RT-PCR)和Western blot实验观察RNAi对hTERT基因的抑制效率;四甲基偶氮唑盐(microculture tetrazolium,MTT)实验检测胃癌细胞增殖情况,利用流式细胞仪检测细胞周期的改变,应用Transwell小室观察胃癌细胞转移能力,划痕实验检测胃癌细胞平面迁移能力。结果胃癌细胞株SGC-7901特异性hTERT基因沉默能够在基因和蛋白水平,特异地抑制hTERT的表达,抑制效率达80%以上,特异性hTERT siRNA能够抑制细胞生长,细胞的增值指数下降(P<0.05),hTERT基因干扰后细胞迁移能力减低(P<0.05)。结论hTERT基因沉默能抑制胃癌细胞增殖、迁移能力。Objective To determine the effect of human telomerase reverse transcriptase （hTERT） knockdown by small interfering RNA （RNA） on the proliferation and migration of gastric cancer SGC-7901 cells. Methods Gastric cancer cell lines SGC-7901 was transfected by hTERT siRNA and ascertained by fluorescence microscope. The inhibitory effect of RNAi on hTERT was observed by real time polymerase chain reaction （RT-PCR） and Western blot. Ceil growth situation, the changes of cell cycle, the cell migration ability and plane migration ability were assessed by microculture tetrazolium （MTT） assay, flow cytometry, Transwell assay and the scratch test respectively. Results Transfection of specific hTERT siRNA into SGC-7901 cells resulted in a specific and highly-efficient declined expression of hTERT （80% ） at mRNA and protein levels, and an inhibition of cell proliferation （P〈0.05）. The transfection of specific hTERT siRNA also led to the decrease of migration ability of SCC 7901（P〈0. 05）. Conclusion HTERT knockdown can inhibit the proliferation, cell tumorigenesis and migartion ability of gastric cancer cells.

关 键 词：人端粒酶逆转录酶 小干扰RNA 胃癌 SGC-7901细胞 

分 类 号：R735.2[医药卫生—肿瘤]