口腔鳞癌细胞中ITGB6基因主要转录调控区域的定位分析  

Site-directed analysis of key transcriptionaI regulatory regions of ITGB6 gene in oral squamous cell carcinoma cells

作　　者：陈锡和[1,2] 邓小玲[3] 尹丽琴[2] 傅玉才[2] 许铭炎[1] 

机构地区：[1]汕头大学医学院第二附属医院口腔科,广东汕头515041 [2]汕头大学医学院细胞衰老实验室,广东汕头515041 [3]汕头大学医学院细胞生物与遗传学教研室,广东汕头515041

出　　处：《癌变．畸变．突变》2013年第3期190-193,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金项目(30900661;81072208);教育部留学回国人员科研启动基金;汕头大学医学院基础与临床科研基金项目

摘　　要：目的:鉴定ITGB6基因在口腔鳞癌细胞中的主要调控区域,为研究基因在口腔鳞癌细胞中的转录调控机制奠定基础。方法:构建含有ITGB6基因转录起始点上游5'端侧翼区不同长度DNA序列的重组pGL2荧光素酶报告基因质粒,转染口腔鳞癌细胞株TCA8113和SAS,利用双荧光素酶报告基因系统检测启动子转录活性,并通过生物信息学方法预测ITGB6基因中潜在的主要转录因子结合位点。结果:获得一系列不同长度ITGB6基因5'侧翼区序列的重组荧光素酶报告基因质粒;当ITGB6基因5'端侧翼片段截短到-187～-35和-35～+27之间时,启动子活性均显著下降;生物信息学分析显示,在-187～-35区域有2个Ets-1的潜在结合位点,而在-35～+27区域有1个IRF-4潜在结合位点。结论:在口腔鳞癌细胞中,ITGB6基因-187^-35和-35^+27区域是主要的转录因子结合区。OBJECTIVE： To identify the key transcriptional regulatory regions in oral squamous cell carcinoma （OSCC） cell lines,and lays the groundwork for investigating the mechanism of ITGB6 transcriptional regulation in OSCC. METHODS：The recombinant pGL2 luciferase reporter constructs containing different lengths of 5＇ -flanking DNA fragments upstream of transcription initiation site of ITGB6 gene were constructed,and were transfected into two OSCC cell lines TCA8113 and SAS. The promoter activities were detected using dual-luciferase reporter assay system and the potential transcription factor binding sites at key transcriptional regulatory region of ITGB6 gene were predicted by bioinformatics method. RESULTS：The recombinant reporter constructs containing different lengths of 5＇ -flanking region of ITGB6 gene were obtained. When the length of ITGB6 5＇ -flanking region was reduced from -187 to -35 and -35 to ＋27,the transcriptional activity decreased significantly. Two potential Ets-1 binding sites and one potential IRF-4 binding site were identified in the region of -187 to ＋27 of ITGB6 gene. CONCLUSION：The -187/-35 and -35/＋27 regions are the two key transcriptional regulatory regions of ITGB6 gene in OSCC cell lines.

关 键 词：口腔鳞癌 ITGB6基因 主要调控区 转录因子 荧光素酶报告基因分析 

分 类 号：R394.3[医药卫生—医学遗传学]