中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (11): 1933-1936.doi: 10.3969/j.issn.1673-8225.2012.11.008

• 组织构建与生物活性因子 tissue construction and bioactive factors • 上一篇    下一篇

转录因子Nkx2.5和GATA4的相互作用*☆

丁建东1,方  翔1,李开如2,姚玉宇1,陶绍玉1,王  点1,马根山1   

  1. 1东南大学附属中大医院心血管内科,江苏省南京市   210009;2安徽医科大学滁州临床学院,安徽省滁州市  239000
  • 收稿日期:2011-08-08 修回日期:2011-11-06 出版日期:2012-03-11
  • 作者简介:丁建东☆,男,1965年生,江苏省盐城市人,汉族,2003年南京医科大学毕业,博士,主任医师,副教授,主要从事先心病的介入治疗及发病机制的研究。dingjiandong@163.com
  • 基金资助:

    受南京市社会发展基金资助项目(200601055)。

Interaction of cardiac transcription factor Nkx2.5 and GATA4

Ding Jian-dong1, Fang Xiang1, Li Kai-ru2, Yao Yu-yu1, Tao Shao-yu1, Wang Dian1, Ma Gen-shan1   

  1. 1Department of Cardiology, the Affiliated Zhongda Hospital of Southeast University, Nanjing  210009, China; 2Chuzhou Clinical College of Anhui Medical University, Chuzhou  239000, Anhui Province, China
  • Received:2011-08-08 Revised:2011-11-06 Online:2012-03-11
  • About author:Ding Jian-dong☆, Doctor, Chief physician, Associate professor, Department of Cardiology, the Affiliated Zhongda Hospital of Southeast University, Nanjing 210009, China dingjiandong@163.com
  • Supported by:

    Nanjing Social Development Fund, No.200601055*

摘要:

背景:心脏发育过程是复杂多种因子相互作用的结果,对转录因子Nkx2.5和GATA4在心脏发育过程中相互作用的研究将有助进一步了解心脏发育过程。
目的:分析心脏转录因子Nkx2.5和GATA4在细胞内的相互作用。
方法:在构建Nkx2.5和GATA4真核表达质粒以及脑钠肽启动子荧光表达质粒的基础上,将真核表达质粒经共转染或单转染COS7细胞,利用免疫共沉淀技术验证Nkx2.5和GATA4在细胞内是否有相互作用,并通过荧光素酶分析技术分析Nkx2.5和GATA4对脑钠肽启动子序列可能存在的协同激活作用。
结果与结论:通过免疫共沉淀实验发现GATA4能将Nkx2.5沉淀下来,相应的Nkx2.5也能将GATA4沉淀下来,说明在COS7细胞中,GATA4能和Nkx2.5相互作用。脑钠肽启动子荧光表达质粒并与Nkx2.5和GATA4共转染COS7细胞,比起单转染Nkx2.5或GATA4质粒,两者共转染更能激活脑钠肽启动子的表达。Nkx2.5和GATA4不仅各自在心脏发育过程中发挥重要作用,两者之间的相互协调作用对于心脏的正常发育过程同样非常重要。

关键词: 转录因子, Nkx2.5, GATA4, 脑钠肽, 先天性心脏病

Abstract:

BACKGROUND: Heart development results from the interaction among various factors. Research on the interaction of Nkx2.5 and GATA4 may help us to further understand the process of heart development.
OBJECTIVE: To research the interaction of cardiac transcription factor Nkx2.5 and GATA4 in vitro.
METHODS: On the basis of the construction of plasmids of pCMV-Myc-GATA4, pCMV-HA-Nkx2.5 and pTal-luc-BNP, plasmids were transfected into COS7 cells. The interaction of Nkx2.5 and GATA4 was identified by co-immunoprecipitation and luciferase analysis. The possible synergistic activation of Nkx2.5 and GATA4 on BNP promoter sequence was analyzed by luciferase. 
RESULTS AND CONCLUSION: Co-immunoprecipitation experiments showed that Nkx2.5 and GATA4 could be precipitated by each other, demonstrating that Nkx2.5 and GATA4 could directly interact with each other in COS7 cells. The pTal-luc-BNP combined with Nkx2.5 and GATA4 were co-transfected into COS7 cells. Compared with single transfected with Nkx2.5 or GATA4 plasmid, co-transfected with Nkx2.5 and GATA4 displayed synergistic transcriptional activation of the BNP promoter. Nkx2.5 and GATA4 play important roles separately in the process of heart development, and the interactions of Nkx2.5 and GATA4 are also vital in normal heart development.

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