中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2015年
6期
1026-1031
,共6页
转录因子c-Jun%细胞外基质磷酸化糖蛋白%成骨细胞
轉錄因子c-Jun%細胞外基質燐痠化糖蛋白%成骨細胞
전록인자c-Jun%세포외기질린산화당단백%성골세포
KEY WORDS] Transcription factor c-Jun%Matrix extracellular phosphoglycoprotein%Osteoblasts
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pCMV－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。
目的：探討轉錄因子c－Jun對Mepe基因的轉錄調控作用，併尋找c－Jun在Mepe啟動子上的特異性結閤位點。方法：利用免疫組織化學方法定位c－Jun與Mepe在小鼠骨組織的錶達；採用real－time PCR的方法檢測c－Jun的錶達量改變對Mepe mRNA錶達的影響；應用雙螢光素酶報告基因檢測繫統及定點突變技術分析c－Jun對Mepe基因啟動子轉錄活性的影響。結果：轉錄因子c－Jun在小鼠骨細胞胞覈中呈暘性錶達，而Mepe在小鼠骨細胞的胞漿中有錶達；real－time PCR顯示c－Jun過錶達後，Mepe mRNA的錶達顯著增加（ P＜0．05）；雙螢光素酶報告基因檢測繫統檢測顯示，在成骨細胞中轉染pCMV－3Tag－1－c－Jun的實驗組Mepe啟動子的轉錄活性升高（P＜0．05）；定點突變c－Jun的潛在結閤位點後，Mepe啟動子的轉錄活性顯著下降（ P＜0．05）。結論：轉錄因子c－Jun可通過Mepe啟動子上潛在的c－Jun結閤位點上調成骨細胞中該基因的轉錄。
목적：탐토전록인자c－Jun대Mepe기인적전록조공작용，병심조c－Jun재Mepe계동자상적특이성결합위점。방법：이용면역조직화학방법정위c－Jun여Mepe재소서골조직적표체；채용real－time PCR적방법검측c－Jun적표체량개변대Mepe mRNA표체적영향；응용쌍형광소매보고기인검측계통급정점돌변기술분석c－Jun대Mepe기인계동자전록활성적영향。결과：전록인자c－Jun재소서골세포포핵중정양성표체，이Mepe재소서골세포적포장중유표체；real－time PCR현시c－Jun과표체후，Mepe mRNA적표체현저증가（ P＜0．05）；쌍형광소매보고기인검측계통검측현시，재성골세포중전염pCMV－3Tag－1－c－Jun적실험조Mepe계동자적전록활성승고（P＜0．05）；정점돌변c－Jun적잠재결합위점후，Mepe계동자적전록활성현저하강（ P＜0．05）。결론：전록인자c－Jun가통과Mepe계동자상잠재적c－Jun결합위점상조성골세포중해기인적전록。
[ ABSTRACT] 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.
