转录因子c-Jun调控成骨细胞Mepe基因表达的研究  

Transcriptional factor c-Jun regulates Mepe gene expression in osteoblasts

作　　者：张娟娟[1] 刘宗霞[1] 孙岩[1] 

机构地区：[1]潍坊医学院口腔学院口腔医学研究所,山东潍坊261053

出　　处：《中国病理生理杂志》2015年第6期1026-1031,共6页Chinese Journal of Pathophysiology

基　　金：山东省自然科学基金资助项目(No.ZR2012HQ036);山东省自然科学基金资助项目(No.ZR2013HQ019);山东省高等学校科技计划项目(No.J12LL51)

摘　　要：目的:探讨转录因子c-Jun对Mepe基因的转录调控作用,并寻找c-Jun在Mepe启动子上的特异性结合位点。方法:利用免疫组织化学方法定位c-Jun与Mepe在小鼠骨组织的表达;采用real-time PCR的方法检测c-Jun的表达量改变对Mepe mRNA表达的影响;应用双萤光素酶报告基因检测系统及定点突变技术分析c-Jun对Mepe基因启动子转录活性的影响。结果:转录因子c-Jun在小鼠骨细胞胞核中呈阳性表达,而Mepe在小鼠骨细胞的胞浆中有表达;real-time PCR显示c-Jun过表达后,Mepe mRNA的表达显著增加(P<0.05);双萤光素酶报告基因检测系统检测显示,在成骨细胞中转染p CMV-3Tag-1-c-Jun的实验组Mepe启动子的转录活性升高(P<0.05);定点突变c-Jun的潜在结合位点后,Mepe启动子的转录活性显著下降(P<0.05)。结论:转录因子c-Jun可通过Mepe启动子上潜在的c-Jun结合位点上调成骨细胞中该基因的转录。AIM：To study the relationship between transcriptional factor c-Jun and Mepe gene expression, and to identify the specific binding site of c-Jun on the promoter of Mepe.METHODS：The expression of c-Jun and Mepe in mouse bone tissue was detected by immunolocalization assay.The mRNA expression of Mepe was determined by real-time PCR when the expression of c-Jun was changed.The techniques of dual luciferase analysis and site-specific mutagenesis were used to measure the effects of c-Jun on the transcriptional activity of Mepe.RESULTS：c-Jun was detected in the nu-cleus of osteocytes, while Mepe was observed in osteocyte cytoplasm.The results of real-time PCR showed that overexpres-sion of c-Jun directly resulted in significantly higher up-regulation of Mepe mRNA.Compared with control group, the tran-scriptional activity of Mepe promoter was increased in osteoblasts which was transfected with pCMV-3Tag-1-c-Jun.Mutation of c-Jun potential binding sites decreased the transcriptional activity of Mepe promoter.CONCLUSION：Mepe gene tran-scription can be up-regulated by c-Jun which binds to the specific sites of Mepe promoter in osteoblasts.

关 键 词：细胞外基质磷酸化糖蛋白 成骨细胞 

分 类 号：R336[医药卫生—人体生理学]