胶质瘤TJ905细胞中微小RNA-125b对靶基因ERBB2的调控作用  

The regulation of miR-125b on its target gene ERBB2 in glioma TJ905 cells

作　　者：万意[1] 蒋栋毅[1] 陈寒春[1] 费喜峰[1] 周强[1] 王之敏[1] 

机构地区：[1]上海交通大学附属苏州九龙医院神经外科,苏州215021

出　　处：《中华实验外科杂志》2013年第9期1800-1802,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30872657）;苏州社会发展项目（SYS201063）

摘　　要：目的 观察微小RNA（miR）-125b在胶质瘤TJ905细胞中与靶基因ERBB2的相互作用.方法 通过生物信息学分析发现miR-125b与ERBB2相互配对的区域,合成miR-125b和对照序列,构建含ERBB2的3＇UTR野生型和突变型的荧光素酶基因,转染TJ905细胞1×108/L,通过双荧光素酶报告基因检测miR-125b对靶基因ERBB2的3＇UTR区的调控作用.转染48 h后提取蛋白,Western blot检测ERBB2的蛋白表达水平.结果 生物信息学方法分析显示miR-125b与ERBB2的3＇UTR区域存在可能的结合位点.与对照组比较miR-125b inhibitor转染组可以上调ERBB2的mRNA水平是对照组的1.7倍,Western blot法检测ERBB2基因是对照组的1.3倍；而miRNA-mimics组ERBB2的mRNA下降（37.19±3.76）％,Western blot法检测ERBB2基因下降（39.87±7.26）％.结论 在胶质瘤TJ905细胞中ERBB2为miR-125b靶基因,miR-125b可以负调节ERBB2的表达.Objective To explore the interaction of microRNA-125b （miR-125b） and its target gene ERBB2 in glioma TJ905 cells.Methods Bioinformatic analysis found that miR-125b and ERBB2 has each other paired regions.We synthesised miR-125b and the control sequence,and constructed containing ERBB2 3＇ UTR of wild-type and mutant luciferase genes,and then transfected into TJ905 cells.We used the double luciferase reporter gene to analyze the function of miR-125b regulation on the 3＇UTR region of the target gene ERBB2.48 h after transfection,protein was extracted,and the ERBB2 protein expression levels were analyzed by Western blotting.Results The bioinformatics analysis showed that miR-125b and ERBB2 3＇UTR region has possible binding sites.Compared with the control group,miR-125binhibitor transfection group can up-regulate the expression of ERBB2 mRNA level was 1.7-fold higher than that of control group,the detection of ERBB2 gene Western blotting was about 1.3-fold higher than that of control group,while group niRNA-mimics ERBB2 mRNA fell about （37.19 ± 3.76） ％,the detection of ERBB2 Western blotting gene decreased by （39.87 ± 7.26）％.miR-125b can decline ERBB2 mRNA and protein levels via experimental verification.Conclusion ERBB2 is the target gene of miR-125b in glioma TJ905 cells,and miR-125b can be a negative regulator to ERBB2 expression.

关 键 词：胶质瘤 ERBB2 微小RNA-125b 

分 类 号：R739.4[医药卫生—肿瘤]