上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (6): 660-.

• 论著（基础研究） • 上一篇    下一篇

Menin调控小鼠胚胎早期发育相关基因的研究

张翠平1, 朱雅欣1,2, 张宏利1, 李文毅1, 吴 铃1, 龙红梅1, 李 果1   

  1. 1.上海交通大学 医学院上海市内分泌代谢病研究所 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；2.上海市黄浦区中心医院内分泌科, 上海 200002
  • 出版日期:2010-06-25 发布日期:2010-06-28
  • 通讯作者: 李 果, 电子信箱: gli_shnfms@yahoo.cn。
  • 作者简介:张翠平（1983—）, 女, 硕士生；电子信箱: magnificient1130@sina.com。
  • 基金资助:

    国家自然科学基金(30771021, 30800537)

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

ZHANG Cui-ping1, ZHU Ya-xin1,2, ZHANG Hong-li1, LI Wen-yi1, WU Ling1, LONG Hong-mei1, Li Guo1   

  1. 1.Shanghai Institute of Endocrine and Metabolic Diseases, Department of Endocrine and Metabolic Diseases, Shanghai Clinical Center for Endocrine and Metabolic Diseases, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200025, China；2.Department of Endocrine and Metabolic Diseases, The Central Hospital of Huangpu District, Shanghai 200002, China
  • Online:2010-06-25 Published:2010-06-28
  • Supported by:

    National Natural Science Foundation of China, 30771021, 30800537

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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。

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

Abstract:

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. Methods Undifferentiated 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. Results It 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). Conclusion Menin may affect mouse embryonic development by several cellular signaling pathways, including Wnt, MAPK, TGF-β and apoptosis signaling pathways.

Key words: menin, embryoid bodies, chromosome immunoprecipitation, DNA microarray, signaling pathway