整合素β6基因在HepG2细胞中主要转录调控区的定位分析  

Site-directed analysis of key transcriptional regulatory regions of human integrin beta6 gene in HepG2 cells

作　　者：许锐锐[1] 韩慧[1] 贺文静[1] 陈耿臻[1] 

机构地区：[1]汕头大学医学院第二附属医院微创医学中心,汕头515000

出　　处：《中华普通外科学文献（电子版）》2013年第2期11-14,共4页Chinese Archives of General Surgery(Electronic Edition)

摘　　要：目的鉴定整合素β6基因在HepG2细胞中的主要转录调控区,为进一步研究αvβ6在肝癌发生发展过程中的转录调控机制奠定基础。方法利用β6基因转录起始位点附近1135 bp基因序列经PCR进行不同长度的缺失扩增,并连接到pGL2-basic载体,构建重组质粒,利用双荧光素酶报告基因分析系统检测启动子的转录活力,并利用Transcription Element Search System预测β6基因主要转录调控区潜在转录因子结合位点。结果获得4个含不同长度β6基因序列的重组质粒;当β6基因在距转录起始位点-458bp截短到-187bp时,转录活力降低约50%,在这个转录调控区存在YY1、AP-1和c-Myb等与肝癌相关的转录因子结合位点。结论在HepG2细胞中,β6基因启动子区的-458bp/-187bp区域是其主要的转录调控区,YY1、AP-1和c-Myb三个转录因子存在该区域,并可能是调控基因转录的重要转录因子,这将有望为肝癌的诊断和治疗提供新的治疗靶点。Objective：To identify key transcriptional regulatory regions of human integrin beta6 （Itgβ6） gene in HepG2 cells, thus to make a preliminary study for its transcriptional regulatory mechanism. Methods：The recombinant plasmids containing different lengths of DNA fragments upstream of translation initiation site of Itgβ6 gene as reporter promoters were constructed by deletion Method, and the promoter activities were detected using dual luciferase reporter assay system. The potential transcription factor binding sites at key transcriptional regulatory region of Itgβ6 gene were predicted by TESS. Results：The recombinant plasmids containing different lengths of Itgβ6 gene were obtained. When the lengths of Itgβ6 gene were reduced from -458 bp to -187 bp, the transcriptional activity was decreased by about 50%. There were YY1、AP-1 and c-Myb transcriptional factor binding sites in the -458 bp to -187 bp regions. Conclusion：The -458 bp/ -187 bp was the key transcriptional regulatory region of Itgβ6 gene in HepG2 cells, YY1、AP-1 and c-Myb are in this region, and may be the important factors for regulating the transcription of Itgβ6 gene.

关 键 词：整合素β6 重组质粒 肝癌 转录因子 

分 类 号：R735.7[医药卫生—肿瘤]