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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2011, Vol. 31 Issue (11): 31-37    
研究报告     
HeLa细胞中INI1通过与E2A作用下调CD117
范彦会1, 韩秋悦2,3, 王雅丽1, 韩春雨1
1. 河北科技大学生物科学与工程学院 石家庄 050018;
2. 石家庄市第九医院 石家庄 050100;
3. 河北医科大学研究生学院 石家庄 050017
INI1 Down-regulates CD117 in HeLa by Interacting with E2A
FAN Yan-hui1, HAN Qiu-yue2,3, WANG Ya-li1, HAN Chun-yu1
1. College of Biological Sciences and Engineering, Hebei University of Science and Technology, Shijiazhuang 050018, China;
2. Ninth Hospital of Shijiazhuang, Shijiazhuang 050100, China;
3. Graduate School, Hebei Medical University, Shijiazhuang 050017, China
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摘要:

目的:已经证明,CD117基因转录启始位点的上游-480bp包括4个E-box和一个GC-box。试验要确定INI1是否通过与E-box结合蛋白的相互作用来调控CD117。方法与结果:实验证明INI1过表达下调 CD117的mRNA水平和荧光素酶的活性,这个酶通过CD117启动子来启动。通过染色质共免疫沉淀反应实验,发现在HeLa细胞未损伤的染色质里面,INI1可以特定的结合CD117的启动子。绘制INI1在CD117启动子区域的结合位点图表明, E-boxes而不是GC-box对于下调CD117启动子的活性是必须的,这一点在Co-IP(共免疫沉淀)实验中被进一步的证实,在这个试验中INI1与E-box结合蛋白E2A在体内相互作用。结论:总的来说,INI1参与调节CD117的表达和细胞增殖。

关键词: INI1CD117E2AHeLa表达    
Abstract:

Objective: It has been demonstrated that the -480 bp upstream of the CD117 gene transcriptional start site contains four E-boxes and one GC-box. It remains to be elucidated, however, whether INI1 regulates CD117 gene by interacting with E-box binding protein. Methods and Results: It is indicated that overexpression of INI1 down-regulates the CD117 mRNA level as well as a luciferase activity driven by CD117 promoter. Through ChIP assays, It shows that INI1 can specifically bind CD117 promoter within intact chromatin of Hela cells. Mapping of the INI1 binding site within the CD117 promoter region shows that the E-boxes but not the GC-box is necessary to down-regulate CD117 promoter activity, and this is further confirmed by a Co-IP experiment, in which INI1 functionally interacts with E-box binding protein E2A in vivo. Conclusion:Taken together, INI1 is involved in the regulation of CD117 expression and cell proliferation.

Key words: INI1    CD117    E2A    Hela    Expression
收稿日期: 2011-07-01 出版日期: 2011-11-25
ZTFLH:  Q786  
通讯作者: 韩春雨     E-mail: hanchunyu@yahoo.com.cn
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引用本文:

范彦会, 韩秋悦, 王雅丽, 韩春雨. HeLa细胞中INI1通过与E2A作用下调CD117[J]. 中国生物工程杂志, 2011, 31(11): 31-37.

FAN Yan-hui, HAN Qiu-yue, WANG Ya-li, HAN Chun-yu. INI1 Down-regulates CD117 in HeLa by Interacting with E2A. China Biotechnology, 2011, 31(11): 31-37.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I11/31


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