Mip1对大鼠心肌细胞凋亡基因Bid转录调控作用的初探  

Study on Transcriptional Regulation Role of Mip1 on Apoptosis-related Gene Bid in H9C2 Cells

作　　者：刘小柳[1] 邓恭华[1] 雷坚[1] 蒋磊[1] 王海云[1] 

机构地区：[1]中南大学湘雅医学院病理生理学系,湖南长沙410078

出　　处：《现代生物医学进展》2009年第7期1230-1232,共3页Progress in Modern Biomedicine

基　　金：国家"973"计划项目(2007CB512000)

摘　　要：目的:研究转录因子Mip1对大鼠心肌细胞H9C2凋亡相关基因Bid的转录调控作用。方法:以H9C2细胞基因组DNA为模板,扩增Bid核心启动子片段,将其克隆入荧光素酶报告基因质粒PGL3-Basic中,构建重组载体,并采用双酶切法、PCR法及基因测序对其进行鉴定。用脂质体转染法将该载体转入Mip1不同程度过表达的H9C2细胞,检测该细胞Bid基因启动子区的转录活性。结果:成功克隆Bid基因启动子区,双酶切、PCR和基因测序均显示PGL3-Basic-Bidpromoter重组载体构建成功。荧光素酶相对活性检测显示在H9C2细胞中,随着Mip1转入的增多,Bid启动子区转录活性逐渐下降。结论:Mip1作为一个新的转录抑制因子,可以下调凋亡基因Bid的转录。BSTRACT Objective： To investigate the transcriptional regulation effects of Mip1 on the apoptosis-related gene Bid in H9C2 cells. Methods： The core promoter regions of Bid gene were amplified by nest PCR using rat genomic DNA as its template, cloned into luciferase reporter gene plasmid PGL3-Basic to construct restructured vector, and then identified by restriction digestion, PCR and gene sequencing. This Restructured vector was transfected into H9C2 cells with over-expressed Mip 1 gene with Lipofectamine, and then the effects of Mip1 on the transcriptional activity of Bid gene promoter regions were detected in H9C2 cells. Results：The Bid gene promoter region was cloned and identified by enzyme digestion. Both PCR and gene sequencing showed that the recombinant vector PGL3-Basic-Bid was successfully constructed. The luciferase activity detection results indicated that the transcription activity of Bid promoter region gradually decreased in H9C2 cells with the transfered Mip1 increased. Conclusions：As a new transcriptional inhibited factor, Mip1 can down-regulate the transcription of apoptosis-related gene Bid.

关 键 词：Mip1 转录调控 BID 荧光素酶报告基因 

分 类 号：Q78[生物学—分子生物学] Q95-3