hsa-mir-373对肿瘤细胞中E-钙黏蛋白表达的影响  

Construction of hsa-mir-373 Expression Plasmid and Its Effect on E-cadherin Protein

作　　者：武卫华[1] 孔璟[1] 叶彬[1] 

机构地区：[1]重庆医科大学分子医学与肿瘤中心基础医学院病原生物学教研室,重庆400016

出　　处：《生物技术通报》2011年第8期167-171,180,共6页Biotechnology Bulletin

基　　金：重庆医科大学校级课题(XBYB2008075)

摘　　要：旨在探讨hsa-mir-373重组真核表达载体对肿瘤细胞中的E-钙黏蛋白(E-cadherin)表达的影响。根据miR-Base数据库中hsa-mir-373序列,设计并构建hsa-mir-373重组表达质粒和对照质粒。采用脂质体转染技术将mir-373重组质粒和阴性对照质粒分别转染BIU87细胞株。采用荧光显微镜观测转染效果。Western blotting检测细胞的E-钙黏蛋白表达量,免疫细胞化学方法检测细胞E-钙粘蛋白的分泌。结果显示,酶切和测序证明hsa-mir-373真核表达载体构建成功。E-钙黏蛋白在转染细胞中表达有明显增加。人工构建的hsa-mir-373载体经转染后可增加肿瘤细胞中E-钙黏蛋白的表达量。The study was to elucidate the effect of hsa-mir-373 on the expression of E-cadherin（E-cad） gene in the tumoural cells.The recombinant mir-373 plasmid and controlled plasmid were designed and synthesized according to the micrornas sequence in miRBase database.The recombinant vectors were transfected into BIU 87 cells by liposome mediated method.The effects of transfection were observated by florescence microscope.The expression of E-cadherin protein was determined by Western blotting and immunocytochemisty.Results showed that Western blotting and immunocytochemisty demonstrated a significant increase of E-cadherin protein levels in cells transfected with mir-373,but not in cells transfected with pGenesil-control,when compared to BIU 87 cells.These data indicated that the expression of E-cadherin can be increased by the recombinated mir-373 plasmid.

关 键 词：has-mir-373 BIU87细胞 E-钙黏蛋白 

分 类 号：R730.2[医药卫生—肿瘤]