microRNA-144~ 451基因簇促进红系分化

基础医学与临床 ›› 2015, Vol. 35 ›› Issue (6): 744-748.

microRNA-144~ 451基因簇促进红系分化

刘晓玲¹,王芳²,赵华路²,余佳²

1. 中国医学科学院基础医学研究所
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

收稿日期:2015-03-19 修回日期:2015-04-15 出版日期:2015-06-05 发布日期:2015-05-27
通讯作者: 余佳 E-mail:j-yu@ibms.pumc.edu.cn;jyu403@gmail.com
基金资助:
国家自然科学基金

MicroRNA-144~ 451 gene cluster promotes erythroid differentiation

Received:2015-03-19 Revised:2015-04-15 Online:2015-06-05 Published:2015-05-27
Contact: Jia YU E-mail:j-yu@ibms.pumc.edu.cn;jyu403@gmail.com

摘要/Abstract

摘要： 目的探索microRNA-144~451基因簇在红系分化过程中的功能及调控机制。方法用real-time PCR法检测microRNA-144和microRNA-451的表达；用ChIP结合real-time PCR的方法检测GATA-1和EKLF在microRNA-144~451基因簇上游的结合及调控；使用microRNA-144和microRNA-451模拟物转染K562细胞，并用联苯胺染色和real-time PCR方法检测K562细胞向红系分化；用Western blot方法检测红系分化抑制因子GATA-2表达。结果 microRNA-144和microRNA-451在K562细胞红系分化过程中呈现表达逐渐上升趋势；转录因子GATA-1和EKLF可以结合在microRNA-144~451基因簇并激活microRNA-144或microRNA-451的表达；在K562细胞中过表达microRNA-144或microRNA-451均可促进hemin诱导的红系分化；另一方面，microRNA-144和microRNA-451可以通过抑制GATA-2的表达促进细胞向红系分化。结论 microRNA-144~451基因簇在红系分化过程中受到GATA-1和EKLF的激活调控，同时通过抑制GATA-2的表达促进红系发育成熟。

关键词: microRNA-144, microRNA-451, GATA-1, EKLF, GATA-2

Abstract: Objective To study the roles of microRNA-144~451 gene cluster during erythroid differentiation of K562 cells. Methods The expression of microRNA-144 and microRNA-451 during K562 erythroid differentiation was detected by quantitative RT-PCR. The occupancy and regulation of transcription factors GATA-1 and EKLF on microRNA-144~451 gene was investigated by chromatin immunoprecipitation (ChIP) and quantitative PCR. K562 cells were transfected with the microRNA-144 or microRNA-451 mimics, respectively. The effects of over-expression of microRNA-144 or microRNA-451 on erythroid differentiation were examined by benzidine staining and quantitative RT-PCR. Western blot was performed to detect the level of erythroid antagonist GATA-2 upon miRNA over-expression. Results microRNA-144 and microRNA-451 were up-regulated during K562 erythroid differentiation. Erythroid transcription factors GATA-1 and EKLF reside and activate microRNA-144~451 gene expression. Over-expression of microRNA-144 or microRNA-451 in K562 cells accelerates its erythroid differentiation. GATA-2 was inhibited by microRNA-144 and microRNA-451 in erythroid cells. Conclusion the microRNA-144~451 gene cluster was activated by GATA-1 and EKLF, and regulated erythroid differentiation of K562 cells via repressing GATA-2.

Key words: microRNA-144, microRNA-451, GATA-1, EKLF, GATA-2

中图分类号:

Q254

刘晓玲王芳赵华路余佳. microRNA-144~ 451基因簇促进红系分化[J]. 基础医学与临床, 2015, 35(6): 744-748.