RNA Interference-Mediated Gene Silencing of Vascular Endothelial Growth Factor  

作　　者：Tiejun Li Jianning Song Kai Kang Zanlan Hu Tongchuan He Bingqiang Zhang Caiquan Zhang 

机构地区：[1]The First Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing, 400016, China [2]The Ninth Hospital of Chongqing, Chongqing, 400700, China [3]Molecular Oncology Laboratory, The University of Chicago Medical Center, Chicago IL 60637, USA

出　　处：《Chinese Journal of Clinical Oncology》2006年第6期408-412,共5页中国肿瘤临床（英文版）

基　　金：This work was supported in part by re-search grants from the National NaturalScience Foundation of China (No.30300298), and the National NaturalScience Foundation of China's JointResearch Fund for Overseas ChineseYoung Scholars Grant (No. 30228026).

摘　　要：OBJECTIVE To inhibit the expression of the vascular endothelial growth factor (VEGF) by RNA interference, and to observe the effect in different cells line. METHODS Using the services of E-RNAi, we designed and constructed two kinds of shRNAs expression vectors which were aimed at the VEGF gene. These vectors were then transfected into HEK293, colon cancer HT29, Hela and HepG2 cells by LipofectamineTM 2000. The level of VEGF mRNA was determined by RT-PCR and Northern blotting and the VEGF expression was examined by immunofluoresence staining. RESULTS The two kinds of VEGF specific shRNAs expression vectors were found to efficiently inhibit the expression of VEGF in HEK293 and HT29 cells by RT-PCR analysis, with inhibition rates of 72% and 42%, respectively; but the inhibition rates were reduced to 28% in Hela cells and 13% in HepG2 cells. Northern blotting showed that the inhibition rates of VEGF mRNA expression were 88% and 89% in HEK293 and HT29 cells, respectively. The inhibition rate of VEGF protein expression in HT29 cells was 73% based on immunofluoresence staining. CONCLUSION The expression of VEGF was inhibited by RNA interference, but differed with various cells lines, showing that RNA interference was cell-line dependent.OBJECTIVE To inhibit the expression of the vascular endothelial growth factor （VEGF） by RNA interference, and to observe the effect in different cells line. METHODS Using the services of E-RNAi, we designed and constructed two kinds of shRNAs expression vectors which were aimed at the VEGF gene. These vectors were then transfected into HEK293, colon cancer HT29, Hela and HepG2 cells by LipofectamineTM 2000. The level of VEGF mRNA was determined by RT-PCR and Northern blotting and the VEGF expression was examined by immunofluoresence staining. RESULTS The two kinds of VEGF specific shRNAs expression vectors were found to efficiently inhibit the expression of VEGF in HEK293 and HT29 cells by RT-PCR analysis, with inhibition rates of 72% and 42%, respectively; but the inhibition rates were reduced to 28% in Hela cells and 13% in HepG2 cells. Northern blotting showed that the inhibition rates of VEGF mRNA expression were 88% and 89% in HEK293 and HT29 cells, respectively. The inhibition rate of VEGF protein expression in HT29 cells was 73% based on immunofluoresence staining. CONCLUSION The expression of VEGF was inhibited by RNA interference, but differed with various cells lines, showing that RNA interference was cell-line dependent.

关 键 词：RNA interference VEGF tumor cells Northern blotting. 

分 类 号：R733[医药卫生—肿瘤]