人Snail基因RNAi慢病毒载体对鼻咽癌5-8F细胞增殖和侵袭的影响  

Lentiviral vector for RNA interference of human Snail gene inhibits proliferation and invasion of nasopharyngeal carcinoma 5-8F cell line

作　　者：陈丽[1] 刘求真[1] 邱际华[1] 焦锋[1] 姚开泰[1] 

机构地区：[1]南方医科大学肿瘤研究所,广州510515

出　　处：《第三军医大学学报》2009年第12期1151-1154,共4页Journal of Third Military Medical University

基　　金：广东省科技计划项目(2007B031515006)~~

摘　　要：目的构建人Snail基因慢病毒干扰载体及观察干扰前后对鼻咽癌5-8F细胞增殖和侵袭的影响。方法设计Snail基因特异性RNAi靶序列,克隆至经双酶切后的plVTHM线性化载体,测序鉴定正确后,用病毒上清感染鼻咽癌细胞5-8F,流式细胞仪分选获得稳定干扰Snail基因的细胞亚系。荧光定量PCR检测mRNA表达情况;MTT法和体外侵袭实验分别测定对细胞增殖和侵袭的影响。结果plVTHM-siSnail慢病毒干扰载体构建正确。荧光定量PCR结果表明分选后的细胞Snail mRNA水平表达显著降低。干扰后的细胞增殖减慢,穿透基膜的细胞数量减少,差异具有统计学意义(P<0.05)。结论plVTHM-siSnail表达质粒可显著下调Snail基因在5-8F中的表达,在一定程度上抑制鼻咽癌细胞的增殖和侵袭。Objective To construct a recombinant lentiviral expression vector for RNA interference （RNAi） of human Snail gene and to study its effects on the proliferation and invasion of nasopharyngeal carcinoma cell line 5-8F. Methods The effective sequence of short hairpin RNAs （shRNA） targeting Snail gene was designed and cloned into the linear pLVTHM vector after enzyme digestion. After confirmation by DNA sequen- cing, 5-8F cells were infected with the viral supernatants. The cells with stable Snail gene knock-down were separated by fluorescence activated cell sorter （FASC）. The expression of Snail mRNA was detected by real time RT-PCR. MTF and cell invasion assay were used to detect the proliferation and invasion of 5-8F cells after plVTHM-siSnail transfection. Results The lentivirus vector plVTHM-siSnail was constructed successfully. The separated 5-8F-plVTHM-siSnail exhibited significant knock-down of Snail mRNA expression. Slower prolif- eration and decreased cells to permeate through the Matrigel were found after plVTHM-siSnail transfection （P 〈 0.05 ）. Conclusion plVTHM-siSnail vector could down-regulate Snail gene expression in human nasopharyngeal carcinoma 5-8 F cells and suppress its invasion and proliferation to some extent.

关 键 词：SNAIL RNA干扰 鼻咽癌 增殖 侵袭 

分 类 号：R394.3[医药卫生—医学遗传学] R73-362[医药卫生—基础医学]