NCOA2核心启动子甲基化分析及其与肾周和皮下脂肪组织差异表达的关系  

NCOA2 core promotor methylation and its differential expression profile in perirenal and subcutaneous fat tissue

作　　者：李艳[1] 孙文星[1] 徐春瑛[1] 褚维伟[1] 谷淑华[1] 陈杰[1] 

机构地区：[1]南京农业大学动物科技学院,江苏南京210095

出　　处：《南京农业大学学报》2014年第4期131-136,共6页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(31272423)

摘　　要：为探讨核受体辅激活蛋白2(NCOA2)基因启动子区甲基化水平与其在肾周脂肪组织及皮下脂肪组织中差异表达的相关性,采用实时荧光定量PCR(qRT-PCR)方法检测NCOA2基因在肾周脂肪组织和皮下脂肪组织中的表达水平;通过构建系列NCOA2基因5'侧翼区缺失表达载体,并采用双荧光素酶报告系统鉴定NCOA2基因核心启动子区;在分析核心启动子区CpG岛的基础上,采用亚硫酸氢盐测序(BSP)法测定NCOA2基因核心启动子区CpG岛甲基化水平。结果表明:肾周脂肪组织中NCOA2基因的mRNA表达水平显著低于皮下脂肪组织;NCOA2基因核心启动子区位于-483^-179 bp;预测发现该区域存在1个CpG岛,BSP法表明该区域内的25个CpG位点在皮下和肾周脂肪组织均为非甲基化状态,无明显组织差异性。结论:肾周脂肪组织和皮下脂肪组织中NCOA2基因表达存在显著差异,而核心启动子区甲基化状态没有参与差异调控。To investigate the relationship between the DNA methylation status of nuclear receptor coactivator 2（NCOA2） and the differential expression of NCOA2,which between in subcutaneous fat tissue and in perirenal fat tissue,the mRNA expression level in perirenal and subcutaneous fat tissue was assessed by quantitative real-time reverse transcription-polymerase chain reaction（qRTPCR）.NCOA2 promoter activity was detected by dual-luciferase reporter system.The DNA methylation status of CpG island in the core promoter region was analyzed by bisulphite sequencing PCR（BSP）.The result showed that NCOA2 mRNA level of in perirenal fat tissue was significantly lower than in subcutaneous fat tissue. NCOA2 promoter activity was detected by dual-luciferase reporter system,which revealed that the from-483 to-179 bp region harbored core promoter of the gene.NCOA2 core promoter was unmethylated in either subcutaneous or perirenal fat tissue.Conclusions： the differential expression of NCOA2 in subcutaneous and perirenal fat tissues had no significant correlation with its core promoter methylation levels.

关 键 词：NCOA2基因 肾周脂肪 皮下脂肪 核心启动子 甲基化 

分 类 号：S813.24[农业科学—畜牧学]