Menin调控小鼠胚胎早期发育相关基因的研究  

Effects of menin on related genes of early development of mouse embryos

作　　者：张翠平[1,2] 朱雅欣[1,2,3] 张宏利[1,2] 李文毅[1,2] 吴铃[1,2] 龙红梅[1,2] 李果[1,2] 

机构地区：[1]上海交通大学医学院上海市内分泌代谢病研究所 [2]上海交通大学瑞金医院内分泌代谢病科上海市内分泌代谢病临床医学中心,上海200025 [3]上海市黄浦区中心医院内分泌科,上海200002

出　　处：《上海交通大学学报（医学版）》2010年第6期660-664,673,共6页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(30771021;30800537)~~

摘　　要：目的研究1型多发性内分泌肿瘤综合征相关基因(Men1)编码的蛋白(menin)对小鼠胚胎早期发育相关基因的调控,并探讨其可能的作用机制。方法以经丝裂霉素C处理的小鼠胚胎成纤维细胞作为滋养层细胞培养未分化的小鼠胚胎干细胞,使其体外分化形成胚胎体。RT-PCR鉴定胚胎体胚胎发育期标志基因表达;染色体免疫共沉淀结合启动子基因芯片技术检测成熟胚胎体中menin调控的下游基因,Real-Time PCR鉴定结果。结果 RT-PCR结果表明胚胎体相当于小鼠胚胎发育的早期器官形成期。基因芯片检测显示,menin能调控8 640个备选基因中的784个;基因分析显示,menin能通过多个信号通路调控下游基因,包括Wnt信号通路的1500003O03Rik、Prkca、Fosl1、Nfatc3、Dvl1;TGF-β信号转导途径的Thbs1、Tgfb3、Bmp4、Smurf2;凋亡信号通路的Casp6、1500003O03Rik、Prkar1a、Ripk1、Traf2、Capn5、Endog、Myd88;MAPK信号通路的Casp6、Arrb2、Fgf15、1500003O03Rik、Map3k4、Map2k1ip1、Tgfb3、Prkca、Traf2、Dusp7、Atf2等。结论 Menin可能通过Wnt、MAPK、TGF-β、凋亡等多种信号通路对小鼠胚胎早期发育相关基因进行调控。Objective To investigate the effects of menin,product of multiple endocrine neoplasia type 1（Men1） gene,on related genes of early development of mouse embryos,and explore its possible mechanism.MethodsUndifferentiated mouse embryonic stem cells were maintained on a feeder layer of Mitomycin C-treated mouse embryonic fibroblasts,and embryoid bodies（EBs） were formed after initiating differentiation.The expression of marker gene in embryonic development period of EBs was identified by RT-PCR.Chromosome immunoprecipitation and DNA microarray were conducted to detect the downstream genes regulated by menin in mouse EBs,and the results were identified by Real-Time PCR.ResultsIt was revealed by RT-PCR that EBs corresponded to the early organogenesis stage of mouse embryonic development.In the microarray containing 8 640 oligonucleotides representing about 8 000 genes in the genome,784 genes were identified as downstream.These genes were then classified into cellular signaling pathways and were identified,including Wnt signaling pathways（1500003O03Rik,Prkca,Fosl1,Nfatc3 and Dvl1）,MAPK signaling pathways（Casp6,Arrb2,Fgf15,1500003O03Rik,Map3k4,Map2k1ip1,Tgfb3,Prkca,Traf2,Dusp7 and Atf2）,TGF-β signaling pathways（Thbs1,Tgfb3,Bmp4 and Smurf2） and apoptosis signaling pathways（Casp6,1500003O03Rik,Prkar1a,Ripk1,Traf2,Capn5,Endog and Myd88）.ConclusionMenin may affect mouse embryonic development by several cellular signaling pathways,including Wnt,MAPK,TGF-β and apoptosis signaling pathways.

关 键 词：MENIN 胚胎体 染色体免疫共沉淀 启动子基因芯片 信号通路 

分 类 号：Q343[生物学—遗传学]