MicroRNA expression dynamics during murine and human erythroid differentiation
MicroRNA expression dynamics during murine and human erythroid differentiation
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DOI：
10.1016/j.exphem.2007.03.014
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发表时间：
2007-07-01
影响因子：
2.6
通讯作者：
Song, Chao-Zhong
中科院分区：
文献类型：
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作者：
Zhan, Mei;Miller, Chris P.;Song, Chao-Zhong
Objective. microRNAs（miRNAs）是一类丰富的小分子非编码RNA，通过序列特异性抑制基因表达来调节细胞的多种功能。我们确定了在红系分化过程中miRNA的表达谱和在红系分化中的假定作用。利用微阵列分析295个miRNAs在其红系分化诱导前后的表达谱。采用白细胞介素活化细胞分选法分离小鼠脾成红细胞。体外诱导人脐血CD 34（+）祖细胞分化。实时荧光定量逆转录聚合酶链反应（Real-time reverse transcriptase polymerase chain reaction，RT-PCR）验证微阵列的结果。将合成的miR-451过表达或敲低的寡核苷酸转染到MEL细胞中。在红系细胞中发现了超过100种miRNAs的表达。其中大多数显示其表达水平随着红系分化的进展而变化。进一步的分析显示，总体miRNA表达水平在红系分化后增加。在所分析的miRNAs中，miR-451在红系成熟期间最显著上调。使用功能获得和功能丧失方法进行的功能研究表明，miR-451与红系成熟相关。在红系细胞分化过程中，miRNA表达发生动态变化，在红系细胞终末分化后，miRNA水平总体增加。miR-451可能在促进红系分化中发挥作用。(c)2007年国际实验血液学学会。爱思唯尔公司出版
Objective. MicroRNAs (miRNAs) are an abundant class of small noncoding RNAs that regulate diverse cellular functions by sequence-specific inhibition of gene expression. We determined miRNA expression profile during erythroid differentiation and putative roles in erythroid differentiation.Methods. The expression profile of 295 miRNAs before and after their erythroid differentiation induction was analyzed using microarray. Fluorescein-activated cell sorting analysis was used to isolate mouse spleen erythroblasts at different differentiation stages. Human cord blood CD34(+) progenitors were differentiated in vitro. Real-time reverse transcriptase polymerase chain reaction was used to confirm the results of miRNA microarray. Synthetic oligonucleotides for miR-451 overexpression or knockdown were transfected into MEL cells.Results. More than 100 miRNAs were found to be expressed in erythroid cells. The majority of them showed changes in their expression levels with progression of erythroid differentiation. Further analysis revealed that overall miRNA expression levels are increased upon erythroid differentiation. Of the miRNAs analyzed, miR-451 was most significantly upregulated during erythroid maturation. Functional studies using gain of function and loss of function approaches showed that miR-451 is associated with erythroid maturation.Conclusions. Dynamic changes in miRNA expression occurred during erythroid differentiation, with an overall increase in the levels of miRNAs upon terminal differentiation of erythroid cells. MiR-451 may play a role in promoting erythroid differentiation. (c) 2007 International Society for Experimental Hematology. Published by Elsevier Inc.