C/EBPα参与小鼠胰岛β细胞囊泡谷氨酰胺转运子2基因表达的调控  

C/EBPα is involved in transcriptional regulation of mVGLUT2 gene expression

作　　者：吴柱国[1] 刘世明[1] 陈心春[2] 

机构地区：[1]广州医学院第二附属医院,广东广州510260 [2]广东医学院附属深圳市第三人民医院,广东深圳518020

出　　处：《中国病理生理杂志》2010年第1期142-145,共4页Chinese Journal of Pathophysiology

摘　　要：目的:探讨转录调节因子CCAAT增强子结合蛋白α(C/EBPα)在小鼠胰岛β细胞株MIN6的囊泡谷氨酰胺转运子2(VGLUT2)基因表达过程中的调控作用。方法:克隆小鼠VLGUT2基因的启动子区,并对其中C/EBPα的结合位点进行了核酸定点突变,以荧光素酶活力方法观察突变后的启动子活性改变情况。通过电泳迁移率实验(EMSA)检测C/EBPα与含有小鼠VGLUT2启动子区核酸序列的核苷核探针的结合能力,以CEBPαsiRNA特异性抑制转录调控因子C/EBPα的基因表达,通过荧光素酶,RT-PCR和Western blotting方法观察抑制C/EBPα基因表达后小鼠VGLUT2的基因表达情况。结果:在对小鼠VGLUT2启动子区C/EBPα结合位点进行核酸定点突变后,小鼠VGLUT2启动子活性下降约50%,EMSA实验表明C/EBPα可与小鼠VGLUT2启动子区结合,当以C/EBPαsiRNA特异性地下调C/EBPα的表达时,小鼠VGLUT2的启动子活性、mRNA和蛋白水平也相应各自下降约60%、40%及45%。结论:C/EBPα参与了小鼠VGLUT2的基因表达调控。AIM： The main purpose of this study is to investigate the regulatory role of C/EBPα in mouse vesicular glutamate transporter 2 （mVGLUT2） gene expression. METHODS： The promoter region of mVGLUT2 was cloned to PGL3 - basic vector. Site - direction mutation was used to identify the CCAAT - enhancer - binding protein α （C/EBPα） binding site. The promoter activity was observed by luciferase system. The binding between C/EBPα protein and mVGLTU2 promoter region was determined by EMSA. The C/EBPα gene expression was inhibited by its specific siR- NA. RESULTS ： mVGLUT2 promoter activity decreased about 50% after mutation of C/EBPα binding site. EMSA showed that C/EBPα protein bound onto mVGLUT2 promoter region. Meanwhile, when C/EBPα gene expression was inhibited by its specific siRNA, mVGLUT2 promoter activity, mRNA level and protein level were decreased about 60%, 40% and 45%, respectively. CONCLUSION： C/EBPα is involved in the regulation of mVGLUT2 gene expression.

关 键 词：小鼠β细胞 启动子 基因表达调控 CCAAT增强子结合蛋白α 囊泡谷氨酰胺转运子2 

分 类 号：R363[医药卫生—病理学]