小鼠胰岛β细胞(Min6)长链非编码RNA Meg3绑定PRC2沉默Hes1表达  

LncRNA Meg3 Silences Hes1 Expression by Binding to PRC2 in Mice Pancreatic β Cells( Min6)

作　　者：胡艳妹[1] 朱亚男[2] 德伟[2] 

机构地区：[1]铜陵职业技术学院医学系,安徽铜陵244000 [2]南京医科大学基础医学院生物化学与分子生物学实验室,江苏南京210029

出　　处：《河南科技大学学报（医学版）》2016年第2期81-83,90,共4页Journal of Henan University of Science & Technology:Medical Science

基　　金：安徽省2015年度高校自然科学研究基金重点项目(No.KJ2015A374);安徽省2015年度高等教育振兴计划重大项目(No.2015zdjy175)

摘　　要：目的细胞水平研究Meg3与PRC2复合物的关系以及对其下游基因的影响。方法实时定量PCR检测Meg3在Min6细胞细胞核和细胞质的定位,采用RIP技术、UCSC Genome Browser、CHIP技术验证Meg3与Ezh2及其下游基因Hes1的关系。结果 qRT-PCR检测Min6中转染Si-Ezh2、Si-Suz12后Hes1基因的表达显著受到抑制,CHIP结果表明Ezh2能直接绑定在Hes1的启动子区域并介导H3K27me3的修饰过程,而敲除Meg3和Ezh2的表达使Ezh2与H3k27的结合能力降低。结论 lncRNAMeg3在小鼠胰岛Min6细胞中能够绑定核心蛋白复合体PRC2,Ezh2可以直接调控下游转录因子基因Hes1的表达,该发现可以进一步丰富和完善胰岛功能的基因调控网络。Objective To explore the relationship of Meg3 and PRC2 complexes as well as its downstream genes in pancreatic βcells （Min6）. Methods qPCR detection Meg3 in localization of Min6 cell nucleus and cytoplasm,we also verified that the relationship of Meg3,Ezh2 as well as its downstream genes Hes1 by RIP,UCSC Genome Browser and CHIP. Results The expression of Hes1 was significantly suppressed by downregulation of Si-Ezh2 and Si-Suz12 using small interfering RNA in Min6 cells.The results of CHIP assays showed that Ezh2 could directly bind to the promoter region of Hes1 and mediate H3K27me3 modification,while transfection of Meg3 and Ezh2 led to reduced Ezh2 and H3K27 binding ability. Conclusion These results indicated that lncRNA Meg3 could bind PRC2 complex in Min6 cells,and Ezh2 could regulate the expression of downstream transcription factor gene Hes1 directly.The discovery would further enrich and improve the gene regulatory network of pancreatic βcells on Meg3 and PRC2 complexes.

关 键 词：长链非编码RNA 胰岛Β细胞 Meg3 PRC2 Hes1表达 

分 类 号：R587.1[医药卫生—内分泌] Q786[医药卫生—内科学]