MicroRNA expression profiling during human cord blood-derived CD34 cell erythropoiesis
MicroRNA expression profiling during human cord blood-derived CD34 cell erythropoiesis
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DOI：
10.1016/j.exphem.2006.12.002
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发表时间：
2007-04-01
影响因子：
2.6
通讯作者：
McNiece, Ian
中科院分区：
文献类型：
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作者：
Choong, Meng Ling;Yang, Henry He;McNiece, Ian
Objective.对衍生自人脐带血（UCB）CD 34细胞和K562细胞的离体分化红系培养物进行微小RNA（miRNA）表达谱分析，以鉴定参与红细胞生成的miRNA。使两种细胞类型经受刺激红细胞分化的生长因子混合物，并定期收获用于小RNA提取。通过阵列杂交鉴定出与未刺激（第0天）细胞相比表达增加或减少至少1.5倍的miRNA。通过对随机选择的miRNAs进行实时定量聚合酶链反应，证实了表达阵列的有效性。对刺激的UCB衍生的CD 34细胞和K-562细胞之间的分层聚类分析和比较揭示了对红系发育和成熟至关重要的miRNA。对脐血来源的CD 34细胞的相关性分析显示，miR-15 b、miR-16、miR-22和miR-185与红系细胞表面抗原（CD 71、CD 36和CD 235 a）的出现和血红蛋白合成具有强正相关性，而miR-28与这些标志物的表达具有负相关性。与普通髓系/红系祖细胞定型相关的签名miRNA（例如，miR-181家族、miR-221、miR-154）、早期红系定型（例如，miR-32、miR-136、miR-137）和成熟（miR-22、miR-28、miR-185）也通过时间相关性分析鉴定。这些miRNAs可能靶向与细胞发育和分化相关的基因。鉴定用于红细胞生成的可能的签名miRNA。需要进一步的实验来确定这些miRNAs在调节红系定型中的作用。(c)2007年国际实验血液学学会。爱思唯尔公司出版
Objective. MicroRNA (miRNA) expression profiling was performed on ex vivo differentiating erythroid cultures derived from human umbilical cord blood (UCB) CD34 cells and K562 cells to identify miRNAs involved in erythropoiesis.Materials and Methods. Both cell types were subjected to growth factor cocktails stimulating erythroid differentiation and were harvested for small RNA extraction at regular intervals. miRNAs with at least a 1.5-fold expression increase or decrease compared to unstimulated (day 0) cells were identified by array hybridization. Validity of the expression array was confirmed by quantitative real-time polymerase chain reaction on randomly selected miRNAs.Results. Hierarchical clustering analysis and comparison between stimulated UCB-derived CD34 cells and K-562 cells revealed miRNAs that are critical for erythroid development and maturation. Correlation analysis on UCB-derived CD34 cells shows that miR-15b, miR-16, miR-22, and miR-185 have strong positive correlation to the appearance of erythroid surface antigens (CD71, CD36, and CD235a) and hemoglobin synthesis, while miR-28 has an inverse relationship to the expression of these markers. Signature miRNAs associated with common myeloid/erythroid progenitor commitment (e.g., miR-181 family, miR-221, miR-154), early erythroid commitment (e.g., miR-32, miR-136, miR-137), and maturation (miR-22, miR-28, miR-185) were also identified by temporal correlation analysis. These miRNAs are predicted to target genes involved in cell development and differentiation.Conclusion. Probable signature miRNAs for erythropoiesis are identified. Further experimentations are needed to define the roles of these miRNAs in regulating erythroid commitment. (c) 2007 International Society for Experimental Hematology. Published by Elsevier Inc.