Objective: To investigate the expression of miR-451 during erythroid differentiation and its correlation with hematological diseases.
Methods: The expression of miR-451 in erythroid differentiation of mouse hematopoietic stem cells (derived from fetal liver) was analyzed by cell culture, flow cytometry, magnetic bead sorting and qRT-PCR. The expression of miR-451 during erythroid differentiation of mouse erythroid leukemia cells (MEL) was analyzed by cell culture and qRT-PCR. The expression of miR-451 in peripheral blood of mice was detected by qRT-PCR, and the expression of miR-451 in fetal liver (14.5 days) was analyzed by microarray. The nucleated erythroid cells from bone marrow of wild type (WT) mice and β-thalassemia (β-thal) mice were sorted by flow cytometry, and the levels of miR-451 and erythroid genes were detected by qRT-PCR. The expression of miR-451 in peripheral blood of patients with clinical hematological diseases was detected by qRT-PCR.
Results: During the differentiation of mouse hematopoietic stem cells (derived from fetal liver) and MEL cells, the expression levels of miR-451 increased gradually. Compared with WT mice, the expression levels of miR-451 in peripheral blood, 14.5-day fetal liver cells and nucleated erythroid cells (sorted from bone marrow) of β-thal mice were significantly increased(P<0.05). Many erythroid differentiation genes in nucleated erythroid cells (sorted from bone marrow) of β-thal mice decreased. Compared with healthy controls, the expression levels of miR-451 was increased in peripheral blood of patients with β-thalassemia and iron deficiency anemia, while the expression levels of miR-451 was decreased in patients with aplastic anemia and myelodysplastic syndrome.
Conclusion: During erythroid differentiation, the expression levels of miR-451 increases gradually. In hematological diseases, the expression levels of miR-451 is different from that of normal controls, which is expected to become an auxiliary diagnostic index for clinical hematological diseases.
题目: MicroRNA-451在红系分化中的表达水平及与血液疾病的相关性研究.
目的: 探讨microRNA-451（miR-451）在红系分化过程及临床血液疾病中的表达水平以及与该疾病的相关性.
方法: 通过细胞培养、流式细胞检测、磁珠分选和实时荧光定量PCR（qRT-PCR）方法分析miR-451在小鼠造血干细胞（胚胎肝来源）向红系定向分化过程中的表达情况；通过细胞培养和qRT-PCR法分析miR-451在小鼠红系白血病细胞（MEL）向红系定向分化过程中的表达情况；采用qRT-PCR法检测小鼠外周血中miR-451表达水平，并通过mRNA芯片分析小鼠14.5 d的胚胎肝细胞中miR-451的表达水平；通过流式细胞仪分选野生型（WT）小鼠和β地中海贫血（β-thal）小鼠骨髓有核红细胞，并采用qRT-PCR法检测两种小鼠骨髓有核红细胞中miR-451和红系相关基因的表达水平；采用qRT-PCR法检测临床血液疾病患者外周血中miR-451的表达水平.
结果: 在小鼠造血干细胞（胚胎肝来源）和MEL细胞向红系定向分化过程中，miR-451的表达水平均逐步升高；与WT小鼠相比，β-thal小鼠外周血、14.5 d胚胎肝细胞以及骨髓有核红细胞中miR-451表达水平均显著增高（P<0.05），且β-thal小鼠骨髓有核红细胞中多种红系分化基因水平均下降（P<0.05）；与健康对照组相比，β地中海贫血患者（携带者）和缺铁性贫血患者外周血中miR-451表达水平增高（P<0.05），再生障碍性贫血患者和骨髓增生异常综合征患者外周血中miR-451表达水平降低（P<0.05）.
结论: 在红系细胞分化过程中，miR-451表达水平逐渐升高；多种血液疾病患者外周血miR-451的表达水平与正常对照组比较有明显差异，有望成为临床血液疾病的辅助诊断指标.
Keywords: correlation; erythroid differentiation; hematological diseases; microRNA-451.