Effects of miR-210-3p on the erythroid differentiation of K562 cells under hypoxia
Effects of miR-210-3p on the erythroid differentiation of K562 cells under hypoxia
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缺氧条件下miR-210-3p对K562细胞红系分化的影响
DOI：
10.3892/mmr.2021.12202
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发表时间：
2021-06
影响因子：
3.4
通讯作者：
Liu Fang
中科院分区：
文献类型：
--
作者：
Caiyan Hu;Yupeng Yan;Chengbing Fu;Jin Ding;Tiantian Li;Shuqiong Wang;Liu Fang
GATA结合蛋白1(GATA-1)是一种重要的造血转录因子，在血小板、嗜酸性粒细胞、肥大细胞和红细胞等血细胞的产生中起重要作用。GATA-1调控常氧条件下microRNA(miRNAs/miRs)参与红系分化。然而，GATA-1的表达和miR-210-3p在低氧条件下红系分化过程中的调控仍不清楚。本研究检测了缺氧条件下K562细胞红系分化模型中GATA-1和miR-210-3p的表达水平，并通过慢病毒转染法检测了GATA-1、miR-210-3p和Smad2对K562细胞红系分化的影响。本研究检测到GATA-1在低氧条件下表达增加。此外，miR-210-3p被鉴定为红系分化的正调控因子，在红系分化过程中和GATA-1低氧过表达实验中均上调。重要的是，在低氧条件下向红系分化的K562细胞模型中，miR-210-3p以GATA-1依赖的方式上调。利用双荧光素酶报告实验，miR-210-3p被确定为GATA-1介导的红细胞生成调节的下游靶点。MiR-210-3p的功能得失分析证实了其在红系分化中的重要性。此外，还发现Smad2可能是miR-210-3p的下游靶基因。生物信息学预测表明，Smad2介导miR-210-3p诱导的红系分化调控。总而言之，本研究为miRNA调节红系分化提供了新的见解。
GATA binding protein 1 (GATA-1) is one of the most important hematopoietic transcription factors in the production of blood cells, such as platelets, eosinophils, mast cells and erythrocytes. GATA-1 regulates the participation of microRNA (miRNAs/miRs) in erythroid differentiation under normoxia. However, GATA-1 expression and the regulation of miR-210-3p in the context of erythroid differentiation under hypoxia remain unknown. The present study examined the expression levels of GATA-1 and miR-210-3p in the model of erythroid differentiation in K562 cells under hypoxia, and determined the effects of GATA-1, miR-210-3p and SMAD2 on erythroid differentiation through lentivirus transfection experiments. The present study detected increased GATA-1 expression under hypoxia. Moreover, miR-210-3p was identified as a positive regulator of erythroid differentiation, which was upregulated both during erythroid differentiation and in GATA-1 overexpression experiments under hypoxia. Importantly, in the K562 cell model of erythroid differentiation under hypoxia, miR-210-3p was upregulated in a GATA-1-dependent manner. Using a double luciferase reporter assay, miR-210-3p was identified as a downstream target of GATA-1-mediated regulation of erythropoiesis. Gain- or loss-of-function analysis of miR-210-3p identified its importance in erythroid differentiation. Furthermore, it was found that SMAD2 may be a downstream target gene for miR-210-3p. Bioinformatics predictions suggested that SMAD2 mediated miR-210-3p-induced regulation of erythroid differentiation. Collectively, the present study provides novel insights into the miRNA regulation of erythroid differentiation.