利用RNAi技术抑制人舌癌Tca-8113细胞增殖的实验  

Inhibition of the expression of tankyrase 1 and celluar proliferation in Tca-8113 tongue cancer cell line by RNA interference

作　　者：赖军 张萍[2] 朱声荣[3] 陶学金[3] 汤国雄[3] 冀予心[3] 

机构地区：[1]广水市第二人民医院口腔科,湖北广水432721 [2]华中科技大学同济医学院附属同济医院分子肿瘤医学中心,湖北武汉430030 [3]华中科技大学同济医学院附属同济医院口腔医学中心,湖北武汉430030

出　　处：《临床口腔医学杂志》2013年第10期579-582,共4页Journal of Clinical Stomatology

基　　金：国家自然科学基金资助项目(30300331)

摘　　要：目的:通过转染可抑制Tca-8113细胞tankyrase 1表达的siRNA,达到抑制Tca-8113细胞增殖的目的。方法:采用体外转录合成方法,合成针对tankyrase 1的siRNA,并进行Cy-3荧光标记,用脂质体转染至Tca-8113细胞,荧光显微镜观察判断转染效果。应用噻唑蓝(MTT)法和流式细胞仪检测siRNA的作用效果;应用Western blot方法鉴定siRNA抑制tankyrase 1表达效果。结果:tankyrase 1 siRNA组与空白对照组、脂质体对照组相比,可诱导Tca-8113细胞tankyrase 1表达下调,Western blot结果分析显示,转染siRNA后可下调tankyrase 1表达量的1/2,并抑制Tca-8113细胞的增殖(P<0.05),同时改变了细胞周期各时相的细胞分布。结论:tankyrase 1 siRNA在体外实验中可沉默目的基因以及抑制Tca-8113细胞增殖。Objective：To investigate the inhibition of cellular proliferation in tongue cancer Tcca-8113 cell lines by RNA interference,and explore the possibility of the tankyrase 1 as a target of gene therapy for tongue cancer. Method： Tankyrase 1 siRNAs were synthesized using SilencerTM siRNA construction kit and with CY-3 fluorescent labeling. SiRNAs were transfected into Tca-8113 cells by LipofectaminTM 2000. The expression of tankyrase 1 was analyzed by western blot. The biological effect of siRNAs on Tca-8113 cell growth was monitored by MTY and flow cytometry and invert microscope. Result：siRNAs are able to downregulate tankyrase 1 expression. A concurrent growth retardation of Tca-8113 cell line was observed in comparison with the control. Conclusion：Inhibition of tankyrase 1 expression with siRNA technique can inhibit the proliferation of Tca-8113 tongue cancer cells. The tankyrase 1 may be a potential target of geue therapy for tongue can- cer.

关 键 词：Tea-8t 13细胞 肿瘤 实验性 端锚聚合酶 

分 类 号：Q813.1[生物学—生物工程]