VEGF siRNA降低人肝癌细胞株SMMC7721的致瘤性  

REDUCTION OF TUMORIGENICITY OF SMMC7721 HEPATOMA CELL LINE BY VEGF siRNA

作　　者：黄建[1] 王海波[1] 葛盛芳[1] 贾仁兵[1] 黄青山[2] 郑新民[1] 

机构地区：[1]上海交通大学医学院生物化学与分子生物学教研室,上海200025 [2]复旦大学遗传学研究所,上海200433

出　　处：《分子细胞生物学报》2007年第5期277-285,共9页Journal of Molecular Cell Biology

基　　金：上海市教育委员会重点学科建设经费资助(ZDXK2001)。~~

摘　　要：本研究应用RNA干扰(RNAi)技术高效抑制VEGF的表达,明显降低了人肝癌细胞株SMMC7721的致瘤性。化学合成针对人VEGF基因的siRNA,体外瞬时转染人肝癌细胞株SMMC7721,RT-PCR和Elisa法检测表明VEGF siRNA实验组与对照组相比,细胞内VEGF mRNA表达下降了76.16%,VEGF分泌蛋白表达则下降了96.28%,而MTT法结果显示VEGF siRNA对SMMC7721细胞增殖没有明显作用。体内实验中,不同时间对荷SMMC7721细胞瘤裸小鼠进行siRNA直接瘤内注射,同时测量瘤体积,最后取瘤块进行组织切片观察并进行分子生物学分析,结果显示,与对照组相比,VEGF siRNA实验组肿瘤体积明显缩小,瘤内出现细胞坏死,同时肿瘤组织中VEGF mRNA和蛋白表达水平均明显降低。In this study the small interfering RNA（siRNA） targeting human VEGF effectively inhibited the expression of VEGF in human hepatoma cell line, SMMC7721, and could dramatically de- crease the tumorigenicity of SMMC7721 s.c. xe tumor. Chemically synthesized siRNA targeting VEGF was transiently transfected into SMMC7721 cells by lipofectamine, RT-PCR and Elisa analysis suggested that the expression of VEGF mRNA and secreted protein in SMMC7721 cells with VEGF siRNA transfection were suppressed with inhibition rates of 76.16% and 96.28% respectively compared with negative control, but the growth rate of SMMC 7721 cells with VEGF siRNA transfection was the same as the control cells. In vivo test, siRNA was injected directly into implanted tumors and the tumors volume were calculated at different time intervaL Result showed that VEGF siRNA greatly inhibited the growth of tumors tissues, which was consistent with decrease of VEGF mRNA and pro- tein compared with control In addition, the VEGF siRNA-treated group exhibited obvious signs of necrosis compared with controL

关 键 词：小干扰RNA VEGF SMMC7721细胞 

分 类 号：R735.7[医药卫生—肿瘤]