rAAV2-EGFP-Slug-siRNA-U6对人胰腺癌血管生成的影响及其机制探讨  

Effects of rAAV2-EGFP-Slug-siRNA-U6 on angiogenesis of human pancreatic adenocarcinoma and its mechanism

作　　者：崔海宁[1] 陈俭云[2] 王正文[1] 张克君[3] 余壮明[1] 王齐全[1] 王明华[1] 朱东明[1] 宋彩霞[1] 张泽米[1] 

机构地区：[1]海南医学院附属医院普外科,海口570102 [2]深圳大学附属罗湖医院(深圳市第五人民医院)普外科,深圳518001 [3]青岛大学医学院附属医院普外二科,青岛517000

出　　处：《中国免疫学杂志》2013年第11期1161-1164,共4页Chinese Journal of Immunology

基　　金：海南省教育厅高等学校科研项目(No.Hjk2010-34)资助

摘　　要：目的:构建并制备携带目的基因的rAAV2-EGFP-Slug-siRNA-U6(Slug-siRNA)腺相关病毒载体,探讨Slug干扰对人胰腺癌细胞株AsPC-1血管生成的影响及其机制。方法:利用以前试验的方法构建重组质粒pSNAV2.0-EGFP-Slug-siRNA-U6。鸡胚绒毛尿囊膜(CAM)实验检测rAAV2-EGFP-Slug-siRNA-U6对细胞血管生成的影响。Western blot及RT-PCR方法鉴定转染前后AsPC-1细胞Slug与VEGF蛋白及mRNA表达。结果:携带编码靶向Slug的小发夹状干扰RNA序列的腺相关病毒载体rAAV2-EGFP-Slug-siRNA-U6成功构建。rAAV2-EGFP-Slug-siRNA-U6抑制细胞血管生成。Slug-siRNA有效抑制AsPC-1细胞Slug与VEGF蛋白及mRNA的表达。结论:rAAV2-EGFP-Slug-siRNA-U6抑制细胞血管生成,其机制可能与干扰Slug及VEGF的表达有关。Objective： To conduct a recombinant adeno-associated virus-2 carrying Slug-siRNA-U6（ rAAV2-Slug-siRNA-U6）,and to study its influence of angiogenesis in human pancreatic adenocarcinoma cell line AsPC-1 and its mechanism. Methods： The targeting sequence of Slug gene which can be effectively silenced in RNA inference was confirmed and the rAAV2-Slug-siRNA-U6 of recombinant adeno-associated virus-2 vector carring Slug-siRNA was constructed in our previous study. Slug-siRNA inhibits angiogenesis by CAM test,significantly. Western blot and RT-PCR methods was used to detect the expression of Slug and VEGF after being infected AsPC-1 cells by Slug-siRNA. Results： The recombinant adeno-associated virus-2 vector carring Slug-siRNA was constructed successfully,Slug-siRNA inhibits angiogenesis by CAM test,significantly. The expression of Slug and VEGF inhibited by Slug-siRNA in human pancreatic adenocarcinoma cell line AsPC-1 using RT-PCR and Western blot. Conclusion： In CAM Test,RNAi-mediated Slug gene with rAAV silencing rAAV2-EGFP-Slug-siRNA-U6 inhibits angiogenesis. The mechanism of inhibiting angiogenesis of rAAV2-EGFP-Slug-siRNA-U6 may be through Slug and VEGF effectively and selectively silencing.

关 键 词：SLUG 小干扰RNA 腺相关病毒 胰腺癌 血管生成 

分 类 号：R735.9[医药卫生—肿瘤]