人胚肾细胞HEK293中miRNA-128-1表达调控机制的初步研究

doi:10.3969/j.issn.1004-616x.2012.03.009

›› 2012, Vol. 24 ›› Issue (3): 205-208.doi: 10.3969/j.issn.1004-616x.2012.03.009

人胚肾细胞HEK293中miRNA-128-1表达调控机制的初步研究

徐可,刘明发,金涛,蒋嵩山,许海雄

中山大学附属汕头市中心医院神经外科，广东汕头 515000

收稿日期:2011-12-21 修回日期:2012-03-25 出版日期:2012-05-30 发布日期:2012-05-30
通讯作者: 许海雄

The mechanism of miRNA-128-1 expression in HEK293 cells

XU Ke，LIU Ming-fa，JIN Tao，JIANG Song-shan，XU Hai-xiong

Department of Neurosurgery, Affiliated Shantou Hospital of Sun

Received:2011-12-21 Revised:2012-03-25 Online:2012-05-30 Published:2012-05-30
Contact: XU Hai-xiong

摘要/Abstract

摘要： 目的：初步研究人胚肾细胞HEK293中miRNA-128-1的表达调控机制，为进一步探讨miRNA-128-1在胶质瘤中的表达调控奠定基础。方法：通过软件UCSC预测miRNA-128-1的启动子区域；运用软件UCSC、CBIL预测可能与miRNA-128-1启动子区域结合的候选转录因子；构建相关质粒，包括pB-miRNA-128-1、pShNF1、pShc-Myc、pShTAF1、pShYY1、pShMAF、pShRELA2、pShRNUX1和pShNFkappaB。然后将pB-miRNA-128-1分别与上述质粒共转染人胚肾细胞HEK293，检测报告基因活性，确定候选转录因子的作用性质。结果：共转染pShc-Myc可显著抑制pB-miRNA-128-1启动子报告基因的活性；共转染pShNFkappaB可显著升高pB-miRNA-128-1启动子报告基因的活性；共转染pShNF1、pShTAF1、pShYY1、pShMAF、pShRELA2或pShRNUX1对pB-miRNA-128-1启动子报告基因的活性未见明显影响。结论：在HEK293细胞miRNA-128-1的表达调控中，c-Myc可能发挥转录激活作用，而NFkappaB可能发挥转录抑制作用。

关键词: miRNA-128-1, 表达调控, 人胚肾细胞

Abstract: OBJECTIVE: To discuss mechanism of miRNA-128-1 expression in HEK293 cells and lay the foundation for further study of miRNA-128-1 regulation in glioma. METHODS：Through analyzing UCSC，we found the probable miRNA-128-1 promoter area. We used UCSC and CBIL to obtain the predicted transcription factors binding on the miRNA-128-1 promoter area. Related plasmids were constructed，including pB-miRNA-128-1 promoter，pShNF1，pShc-Myc，pShTAF1，pShYY1，pShMAF，pShRELA2，pShRNUX1 and pShNFkappaB. Cotransfecting pB-miRNA-128-1 promoter and pShRNA into HEK293 cells，we analyzed luciferase activity to identify probable transcription factors which could regulate miRNA-128-1 promoter activity. RESULTS：pShc-Myc suppressed the activity of pB-miRNA-128-1 promoter. pShNFkappaB increased the activity of pB-miRNA-128-1 promoter dramatically. pShNF1，pShTAF1，pShYY1，pShMAF，pShRELA2 or pShRNUX1 showed no obvious influence on the expression of pB-miRNA-128-1. CONCLUSION：In HEK 293 cells，c-Myc upregulated whilst NFkappaB downregulated the promoter activity of miRNA-128-1.

Key words: miRNA-128-1, regulation of expression, HEK293 cells

中图分类号:

Q768
R741

徐可,刘明发,金涛,蒋嵩山,许海雄. 人胚肾细胞HEK293中miRNA-128-1表达调控机制的初步研究[J]. , 2012, 24(3): 205-208.

XU Ke，LIU Ming-fa，JIN Tao，JIANG Song-shan，XU Hai-xiong. The mechanism of miRNA-128-1 expression in HEK293 cells[J]. , 2012, 24(3): 205-208.

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人胚肾细胞HEK293中miRNA-128-1表达调控机制的初步研究

The mechanism of miRNA-128-1 expression in HEK293 cells

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