microRNA-144～451基因簇促进红系分化  

MicroRNA-144～451 gene cluster promotes erythroid differentiation

作　　者：刘晓玲[1] 王芳[1] 赵华路[1] 余佳[1] 

机构地区：[1]中国医学科学院基础医学研究所医学分子生物学国家重点实验室,北京100005

出　　处：《基础医学与临床》2015年第6期744-748,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(31471227)

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-I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.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 RTPCR.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.Conclusions The microRNA-144 ～451 gene cluster is activated by GATA-1 and EKLF and then regulates erythroid differentiation of K562 cells via repressing GATA-2.

关 键 词：microRNA-144 microRNA-451 GATA-1 EKLF GATA-2 

分 类 号：Q78[生物学—分子生物学]