EKLF转录激活miR-96在红系分化过程中表达

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (4): 369-374.

EKLF转录激活miR-96在红系分化过程中表达

蒋凤兵¹,余佳²,施琼³,阎文婷¹,朱勇⁴,罗敏⁵,王斌⁶,马艳妮⁴

1. 重庆医科大学基础医学院
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. 重庆医科大学
4. 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
5. 重庆医科大学检验医学院临床检验诊断学教育部重点实验室
6. 山西医科大学第一附属医院普外科

收稿日期:2011-12-29 修回日期:2012-02-23 出版日期:2012-04-05 发布日期:2012-03-21
通讯作者: 施琼 E-mail:anniesq8718@yahoo.com.cn
基金资助:
国家自然科学基金;重庆市自然科学基金

EKLF activates the expression of miR-96 during erythroid differentiation

Received:2011-12-29 Revised:2012-02-23 Online:2012-04-05 Published:2012-03-21
Contact: Qiong SHI E-mail:anniesq8718@yahoo.com.cn
Supported by:
;Natural Science Foundation Project of CQ CSTC

摘要/Abstract

摘要： 摘要：目的研究红系分化过程中，红系分化特异转录因子EKLF对miR-96的表达调控。方法：氯化高铁血红素（Hemin）诱导人K562细胞（人红白血病细胞系）向红系分化，Real-time PCR检测miR-96表达。生物信息学分析miR-96转录起始点上游可能的EKLF结合位点。并经染色质免疫共沉淀(ChIP)与双荧光报告基因实验验证EKLF与这些位点结合的生物学意义。结果 Hemin诱导K562细胞向红系分化过程中，miR-96表达显著升高，且在48h表达最高(3.94156±0.63995, p<0.05)。生物信息学分析显示miR-96转录起始位点上游2.0kb内含有多个EKLF结合位点。经ChIP验证这些位点有EKLF的结合，且EKLF与这些位点结合能募集RNA聚合酶II（Pol II）结合，起始转录。进一步的双荧光报告基因实验也证明EKLF对miR-96转录起始点上游序列具有转录激活作用。结论EKLF转录激活miR-96在红系分化过程中的表达。

关键词: 红系分化, EKLF, miR-96, 染色质免疫共沉淀

Abstract: Abstract：Objective To study the regulation of Erythroid Kruppel-like factor (ELKF) on the expression of miR-96 in Erythropoiesis. Method K562 cells were induced to erythroid differentiation by Hemin. The expression of miR-96 during erythroid differentiation was detected by real-time PCR. Bioinformatic analysis was used to search for binding sites for EKLF within 2kb upstream sequence of transcriptional start site (TSS) of miR-96. Biology function of these sites were detected by ChIP-PCR combined with Dual-Luciferase Reporter Assay. Result miR-96 expression was increased during erythroid differentiation of k562 cells induced by Hemin and peaked at 48h (3.94156±0.63995, p<0.05). Several binding sites within 2kb upstream sequence of TSS of miR-96, occupied with EKLF can recruit RNA polymerase II (Pol II) and promote the expression of downstream genes. Conclusion EKLF activates the expression of miR-96 in Erythropoiesis directly.

Key words: Erythroid differentiation, EKLF, miR-96, ChIP

蒋凤兵余佳施琼阎文婷朱勇罗敏王斌马艳妮. EKLF转录激活miR-96在红系分化过程中表达[J]. 基础医学与临床, 2012, 32(4): 369-374.

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