中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (15): 2730-2734.doi: 10.3969/j.issn.1673-8225.2011.15.017

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

全反式维甲酸诱导小鼠碱性磷酸酶基因启动子区染色体重构

陈  迪，孙奋勇   

  1. 暨南大学生命科学技术学院生物工程研究所，广东省广州市 510632
  • 收稿日期:2010-12-31 修回日期:2011-02-25 出版日期:2011-04-09 发布日期:2013-11-06
  • 通讯作者: 孙奋勇，医学博士，教授，暨南大学生命科学技术学院生物工程研究所，广东省广州市 510632 sunfenyong@ 263.net
  • 作者简介:陈迪★，女，1983年生，河北省定兴县人，满族，暨南大学在读硕士，主要从事基因工程制药方面的研究。 chendi1108@ 126.com
  • 基金资助:

    国家自然科学基金项目(81071524)资助。

All-trans retinoic acid induces chromatin remodeling at the promoter of mouse alkaline phosphatase

Chen Di, Sun Fen-yong   

  1. Institute of Bioengineering, College of Life Science and Technology, Jinan University, Guangzhou  510632, Guangdong Province, China
  • Received:2010-12-31 Revised:2011-02-25 Online:2011-04-09 Published:2013-11-06
  • Contact: Sun Fen-yong, Doctor, Professor, Institute of Bioengineering, College of Life Science and Technology, Jinan University, Guangzhou 510632, Guangdong Province, China sunfenyong@263.net
  • About author:Chen Di★, Studying for master’s degree, Institute of Bioengineering, College of Life Science and Technology, Jinan University, Guangzhou 510632, Guangdong Province, China chendi1108@126. com
  • Supported by:

    the National Natural Science Foundation of China, No. 81071524*

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摘要：

背景：碱性磷酸酶基因是成骨细胞分化和骨形成的重要标志。在C3H10T1/2细胞中，全反式维甲酸可通过核受体上调小鼠碱性磷酸酶的表达，与MAPK通路无关。
目的：从染色体结构调控方面揭示全反式维甲酸上调碱性磷酸酶表达的分子机制。
方法：10-6 mol/L全反式维甲酸处理C3H10T1/2细胞0，1，6，12 h，DNA酶Ⅰ超敏感实验确定全反式维甲酸调控区域的位置，染色质免疫共沉淀实验检验全反式维甲酸处理细胞后一系列转录相关因子与全反式维甲酸调控区域结合的量效关系以及时相分布。
结果与结论：DNA聚合酶Ⅰ超敏感实验表明，小鼠碱性磷酸酶启动子转录起始位点上游约520 bp附近为潜在的全反式维甲酸调控区域；染色质免疫共沉淀实验表明，全反式维甲酸对小鼠碱性磷酸酶的上调作用是通过一系列转录相关因子的时序性共同作用来实现。表明全反式维甲酸诱导小鼠碱性磷酸酶基因转录的过程中伴随着染色质重构和组蛋白的修饰作用。

关键词: 碱性磷酸酶启动子, 全反式维甲酸, 小鼠, C3H10T1/2, 染色体重构, 组氨酸修饰

Abstract:

BACKGROUND: Alkaline phosphatase (ALP) is an important biomarker of proliferation and differentiation in osteoblasts. All-trans retinoic acid (ATRA) up-regulates ALP expression in C3H10T1/2 cells mediated by retinoic acid receptors (RAR) and is irrelevant to the MAPK pathway.
OBJECTIVE: To reveal the mechanism of ATRA induced ALP up-regulation from chromosome structure.
METHODS: C3H10T1/2 were treated with ATRA (10-6 mol/L) for 0, 1, 6 and 12 hours, and then DNaseⅠhypersensitivity analysis was performed to identify the location of the regulatory regions of ATRA. ChIP assays were performed to analyzed transcription factors bound to regulatory regions of ATRA.
RESULTS AND CONCLUSION: DNaseⅠhypersensitivity analysis identified the location of regulatory regions of ATRA which exists in the 520 bp region of ALP promoter. ChIP assays proved that ALP up-regulated by ATRA is accompanied by a series of transcription factors binding to ALP promoter. ALP up-regulated by ATRA is accompanied by chromatin remodeling and Histone modification.

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