微小RNA-223-3p通过调节MYH10基因促进巨核细胞多倍体化

发布时间：2018-01-03 07:26

  本文关键词：微小RNA-223-3p通过调节MYH10基因促进巨核细胞多倍体化　出处：《军事医学》2017年07期 　论文类型：期刊论文

  更多相关文章： miR--p MYH 巨核细胞 聚合酶链反应 流式细胞术

【摘要】：目的探讨微小RNA-223-3p(miR-223-3p)对巨核细胞分化和成熟的影响,并初步探索其中可能的机制。方法通过实时定量PCR检测巨核细胞分化过程中miR-223-3p的内源性表达变化趋势,而后外源调节miR-223-3p在细胞系中的表达量,并通过流式细胞术检测其对于巨核细胞分化和成熟的影响,运用生物信息学分析,找到其发挥相关生物学作用的靶基因MYH10,并通过实时定量PCR、荧光素酶、流式细胞术验证MYH10是miR-223-3p的靶基因。结果内源性miR-223-3p随着巨核细胞的分化成熟表达量增加,在K562和Meg-01细胞系中转染miR-223-3p mimics后可升高巨核细胞相关表面标志CD41和CD61的阳性率,同时显著促进多倍体的形成,MYH10的基因表达随miR-223-3p表达升高而下降,通过双荧光素酶报告基因技术验证MYH10基因是miR-223-3p的靶基因,进一步抑制MYH10基因表达后,可促进巨核细胞多倍体化。结论 miR-223-3p可通过调控MYH10的表达调节巨核细胞的成熟。
[Abstract]:Objective to investigate the effects of microRNA-223-3pmmiR-223-3p) on the differentiation and maturation of megakaryocytes. Methods the endogenous expression of miR-223-3p in megakaryocyte differentiation was detected by real-time quantitative PCR. Then exogenous regulation of miR-223-3p expression in the cell line, and flow cytometry to detect its effect on megakaryocyte differentiation and maturation, bioinformatics analysis. The target gene MYH10 was found to play a related biological role, and real-time quantification of PCR-, luciferase. Flow cytometry confirmed that MYH10 was the target gene of miR-223-3p. Results the expression of endogenous miR-223-3p increased with the differentiation and maturation of megakaryocytes. Transfection of miR-223-3p mimics into K562 and Meg-01 cells increased the positive rate of CD41 and CD61. At the same time, the gene expression of MYH10 decreased with the increase of miR-223-3p expression. Double luciferase reporter gene technique was used to confirm that MYH10 gene is the target gene of miR-223-3p, and further inhibit the expression of MYH10 gene. Conclusion miR-223-3p can regulate the maturation of megakaryocyte by regulating the expression of MYH10.
【作者单位】： 南方医科大学;军事医学科学院野战输血研究所干细胞与再生医学研究室;华南生物医药研究院;
【基金】：国家863计划资助项目(2013AA020107) 国家自然科学基金资助项目(81301132) 北京市自然科学基金资助项目(7152108) 广州市健康医疗协同创新重大专项资助项目(201508020257,201400000003-4)
【分类号】：R3416
【正文快照】： 血小板是由骨髓中巨核细胞生成的具有凝血止血功能的小块胞质,其在人体外周血中的数量如50×109/L,会导致机体出血倾向,当血小板严重减少时,将威胁到人体的生命。目前临床上主要采用机采血小板输注来维持血小板数量,而机采血小板依赖于献血者,且由于献血时间长,舒适度低,血小，

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