基础医学与临床 ›› 2014, Vol. 34 ›› Issue (7): 896-903.

• 研究论文 • 上一篇    下一篇

miR-29a和miR-142-3p促进髓系分化的基因调控网络

董贺1,马艳妮1,董磊2,赵华路2,张俊武3,王芳2,余佳2   

  1. 1. 中国医学科学院 基础医学研究院 北京协和医学院 基础学院 医学分子生物学国家重点实验室
    2. 中国医学科学院基础医学研究所北京协和医学院基础学院
    3. 中国医学科学院基础医学研究所
  • 收稿日期:2014-02-28 修回日期:2014-04-23 出版日期:2014-07-05 发布日期:2014-06-24
  • 通讯作者: 余佳 E-mail:j-yu@ibms.pumc.edu.cn;jyu403@gmail.com
  • 基金资助:
    科技部重大科学问题导向项目

The regulatory network in myeloid differentiation promoted by miR-29a and miR-142-3p

  • Received:2014-02-28 Revised:2014-04-23 Online:2014-07-05 Published:2014-06-24
  • Contact: Jia YU E-mail:j-yu@ibms.pumc.edu.cn;jyu403@gmail.com

摘要: 目的 深入了解miR-29a/miR-142-3p参与促进髓系分化的调控网络。方法 首先实现miR-29a和miR-142-3p在HL-60细胞中的过表达,同时使用PMA以及ATRA分别诱导HL-60细胞向单核及粒系分化。May-Grünwald-Giemsa染色方法观察核形态变化,Real-time PCR检测细胞内CD11和CD14的表达。mRNA表达谱芯片分析获得表达模式相似的基因,利用DAVID和GeneMANIA进行GO categories 和信号通路归类分析。PicTar和TargetScan进行靶基因预测,双荧光报告基因实验确定靶基因的真实性。结果 miR-29a/miR-142-3p过表达细胞与PMA/ATRA诱导分化细胞在整体基因表达模式上有较高的相似度,而表达模式相似的基因大多与细胞分化相关,且在GO分析中显著富集。同时,ZBTB5、CaMKK2、SUV420H2、TRIB2和CANX被确定为miR-29a新的靶基因。结论 miR-29a/miR-142-3p通过调节其靶基因活性,引发整个基因表达调控网络的改变,使之趋向于髓系分化过程中的基因表达变化,从而促进髓系分化进程。

关键词: miR-29a, miR-142-3p, 髓系分化, 基因表达, GO分析

Abstract: Objective To understand the regulatory network of myeloid differentiation promoted by miR-29a/miR-142-3p deeply. Methods Firstly, miR-29a and miR-142-3p were overexpressed in HL-60 cells meanwhile HL-60 cells were also induced towards monocyte and granulocyte differentiation using PMA and ATRA respectively. Used May- Grünwald-Giemsa staining to observe the nuclear morphometry change and utilized Real time PCR to detect the expression of CD11 and CD14 in cells. Genes which possessed similar expression pattern were obtained by microarray and analyzed for their similarities. Analyzed by GO and signal pathway clustering using DAVID and GeneMANIA. Utilizing PicTar and TargetScan, predicted new targets of miR-29a which were also verified by double-luciferase reporter assay. Results Many differential expressed genes during ATRA/PMA induced myeloid differentiation or in the cells with miR-29a/miR-142-3p overexpression were similar. Significantly enriched genes in GO analysis which also possessed similar expression patterns were almost associated with cell differentiation. Conclusion miR-29a and miR-142-3p regulated the activity of their targets, triggered global gene expression change to make it closer to that during myeloid differentiation and then promoted myeloid differentiation. ZBTB5, CaMKK2, SUV420H2, TRIB2 and CANX may play important roles in myeloid differentiation promoted by miR-29a.

Key words: miR-29a, miR-142-3p, myeloid differentiation, gene expression, GO analysis

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