中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (23): 4259-4262.doi: 10.3969/j.issn.1673-8225.2012.23.016

• 干细胞转基因表达 transgenic expression in stem cells • 上一篇    下一篇

干细胞调控因子神经系统多梳蛋白1的全基因组范围结合靶点分析

李 辉1，刘 媛2，龚燕华2，3   

  1. 武警后勤学院， 1组织学与胚胎学教研室，3生物化学与分子生物学教研室，天津市 300162；2中国医学科学院基础医学研究所，北京市 100005
  • 收稿日期:2012-03-12 修回日期:2012-03-31 出版日期:2012-06-03 发布日期:2013-11-06
  • 通讯作者: 龚燕华，博士，副教授，中国医学科学院基础所，北京市 100005；武警后勤学院生物化学与分子生物学教研室，天津市 300162 bigchock@vip.sina.com
  • 作者简介:李辉★，女，1977年生，湖北省襄阳市人，汉族，2004年华中科技大学同济医学院毕业，硕士，讲师，主要从事组织胚胎学与神经生物学研究。 greylh@163.com
  • 基金资助:

    国家自然科学基金资助项目(31070929)；武警后勤学院博士启动金资助项目(WYB201104)。

Whole genome binding sites of stem cell regulator nervous system polycomb 1

Li Hui1, Liu Yuan2, Gong Yan-hua2,3   

  1. 1Department of Histology and Embryology, 3Department of Biochemistry and Molecular Biology, Medical College of Chinese People's Armed Police Force, Tianjin 300162, China; 2Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Beijing 100005, China
  • Received:2012-03-12 Revised:2012-03-31 Online:2012-06-03 Published:2013-11-06
  • Contact: Gong Yan-hua, M.D., Associate professor, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Beijing 100005, China; Department of Biochemistry and Molecular Biology, Medical College of Chinese People's Armed Police Force, Tianjin 300162, China bigchock@vip.sina.com
  • About author:Li Hui★, Master, Lecturer, Department of Histology and Embryology, Medical College of Chinese People's Armed Police Force, Tianjin 300162, China greylh@163.com

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摘要：

背景：在胚胎发育早期神经系统高表达的多梳蛋白家族成员神经系统多梳蛋白1具有明确转录抑制作用。最近研究表明，神经系统多梳蛋白1可能是一个有别于同源基因Bmi1的新的神经干细胞调控因子。
目的：检测体外神经细胞分化模型小鼠畸胎瘤P19细胞中神经发育相关表观遗传调控因子神经系统多梳蛋白1在基因组上的结合靶点。
方法：扩增未分化状态的小鼠畸胎瘤P19细胞系(内源性高表达神经系统多梳蛋白1基因)，使用兔源抗神经系统多梳蛋白1多克隆抗体进行P19细胞的染色质免疫沉淀实验，并结合采用高通量芯片检测技术(ChIP-on-chip)检测捕获靶基因序列，最后利用生物信息学在线软件分析神经系统多梳蛋白1结合靶点临近基因功能的分类特点。
结果与结论：筛选出约1 280个神经系统多梳蛋白1结合靶点，靶点临近或所在基因的功能主要与分化、发育、转录及其调节、神经发生等密切相关。提示全基因组结合靶点分析可有效揭示神经系统多梳蛋白1基因在体外神经干细胞分化模型P19细胞中结合并可能参与调控的靶基因谱的分布特点，有利于下一步深入开展干细胞调控相关因子神经系统多梳蛋白1的下游信号通路研究。

关键词: 神经系统多梳蛋白1, 染色质免疫沉淀, 全基因组, 结合靶点分析, 干细胞调控因子

Abstract:

BACKGROUND: Polycomb group member nervous system polycomb 1 (NSPc1) is a transcription repressor which is highly expressed in the embryo nervous system. Recent research has shown that NSPc1 should be a new regulator of neural stem cell as well as its homologue Bmi1.
OBJECTIVE: To identify the whole genome binding sites of stem cell regulator NSPc1 in the P19 neural differentiation model (mouse embryonal carcinoma cells).
METHODS: Ex vivo expanded undifferentiated P19 cells, which have high expression of endogenous NSPc1 gene, were applied to chromatin immunoprecipitation (ChIP) analysis by using rabbit anti-NSPc1 polyclonal antibody. ChIP-on-chip, a high throughput method of screening the binding targets of NSPc1, was used within these P19 cells. Finally, the functional features of targets were analyzed with bio-informatics methods.
RESULTS AND CONCLUSION: Nearly 1 280 NSPc1 target genes were screened out. Gene Ontology terms associated with "differentiation, development, transcription and its regulation, neurogenesis" were of significantly higher frequency with NSPc1 target genes than their random frequency across the entire genome. Whole genome binding sites analysis is able to indicate the target gene profile of NSPc1 in P19 neural differentiation model, which is very helpful to further investigate the downstream signaling pathway of the new stem cell regulator NSPc1.

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