不同细胞系中RNA干扰抑制血管内皮生长因子表达  

Inhibition of Vascular Endothelial Growth Factor by RNA Interference in Different Cells Lines

作　　者：李铁军[1] 康楷[2] 宋建宁[3] 胡瓒斓[3] 张秉强[1] 张才全[1] 

机构地区：[1]重庆医科大学附属第一医院,重庆400016 [2]重庆医科大学附属第二医院,重庆400010 [3]重庆市第九人民医院,重庆400700

出　　处：《生物技术通报》2007年第1期108-112,共5页Biotechnology Bulletin

基　　金：国家杰出青年基金(30225026);国家青年科学基金(30300298)

摘　　要：目的通过RNA干扰技术抑制血管内皮生长因子(VEGF)表达,并观察在不同细胞系中,RNA干扰作用强度的变化。方法将VEGF基因作为RNA干扰的靶区,通过E-RNAi网上提供的服务,设计两个特异的RNA干扰序列,将其装入含U6启动子的载体上,构建成抗VEGF基因的小发夹样RNA(shRNA)表达载体,再转染人胚肾细胞HEK293、结肠癌细胞HT29、宫颈癌细胞Hela和肝癌细胞HepG2,通过RT-PCR观察VEGF表达受抑的程度及在不同细胞系中RNA干扰作用强度的变化。结果成功构建了两种抗VEGF基因的shRNA表达载体,发现其在HEK293和HT29细胞系中,能明显抑制VEGF基因的表达,抑制率分别为72%和42%;但在Hela细胞中,抑制作用明显减低至28%;在HepG2细胞中抑制作用更弱,仅为13%。结论针对VEGF基因的shRNA表达载体能够明显抑制VEGF基因的表达,但在不同细胞系中的作用强度有明显差别,提示RNA干扰作用存在明显的细胞系选择性。Objective To inhibit the expression of Vascular Endothelial Growth Factor （VEGF）by RNA interference,and observe their vary in different cells line. Methods Followed the service of E-RNAi,we designed and construct two kinds of shRNA expression vectors which aimed VEGF gene,then transfected them into HEK293,colon cancer HT29,Hela and HepG2 cells by lipofectamineTM 2000. The level of VEGF mRNA was investigated by RT- PCR. Results We got two kinds of VEGF specific shRNA expression vectors,and found they could efficiently inhibit the expression of VEGF in HEK293 and HT29,the inhibition rate were 72% and 42%,respectively;but the inhibition rate was cut down to 28% in Hela and 13% in HepG2 cells lines. Conclusion The expression of VEGF could be inhibited by RNA interference,but differed with the change of cells lines,showed that RNA interference was cells lines dependent.

关 键 词：RNA干扰 DICER 小发夹样RNA(shRNA) 

分 类 号：Q78[生物学—分子生物学]