MicroRNA 16 enhances differentiation of human bone marrow mesenchymal stem cells in a cardiac niche toward myogenic phenotypes in vitro
MicroRNA 16 enhances differentiation of human bone marrow mesenchymal stem cells in a cardiac niche toward myogenic phenotypes in vitro
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MicroRNA 16 增强心脏微环境中人骨髓间充质干细胞向体外肌源表型的分化
DOI：
10.1016/j.lfs.2012.05.011
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发表时间：
2012-06-27
期刊：
影响因子：
6.1
通讯作者：
Shan, Zhi-Xin
中科院分区：
文献类型：
--
作者：
Liu, Ju-Li;Jiang, Li;Shan, Zhi-Xin
目的：微小RNA 16（miR-16）的上调有助于人骨髓间充质干细胞（hMSCs）在心脏生态位中向肌源性表型分化，本研究旨在确定miR-16在此过程中的作用。主要方法：hMSCs和新生大鼠心室肌细胞在两个腔室中间接共培养，建立心脏生态位 微环境（利基）。通过 miRNA 微阵列检测心脏生态位诱导的 hMSC 中的 miRNA 表达谱。在不同处理的 hMSC 中测定心脏标志物表达和细胞周期分析。采用实时定量 PCR 和 Western blot 来鉴定 mRNA、成熟 miRNA 和目的蛋白的表达。主要发现：心脏微环境诱导后，hMSC 中显示出 miRNA 失调。 miR-16 在心脏微环境诱导的 hMSC 中上调。 miR-16 的过表达显着增加 hMSC 细胞周期的 G1 期停滞，并增强心脏标志基因的表达，包括 GATA4、NK2-5、MEF2C 和 TNNI3。分化诱导因子 3 (DIF-3) 是一种 G0/G1 细胞周期阻滞化合物，用于诱导心脏微环境诱导的 hMSC 中的 G1 期阻滞，并且在 DIF-3 处理的 hMSC 中心脏标志物基因的表达上调。 hMSCs 中 CCND1、CCND2 和 CDK6 的表达被 miR-16 抑制。 CDK6、CCND1 或 CCND2 敲低导致 hMSC 中的 G1 期停滞，并上调心脏生态位中 hMSC 中的心脏标志物基因表达。意义：miR-16 增强 hMSC 中的 G1 期停滞，有助于 hMSC 在心脏生态位中向肌源性表型分化。该机制为在基于 hMSC 的移植治疗严重心脏病之前对 hMSC 进行预修饰提供了一种新策略。 Crown 版权所有 (c) 2012 由 Elsevier Inc. 出版。保留所有权利。
Aim: Upregulation of microRNA 16 (miR-16) contributed to the differentiation of human bone marrow mesenchymal stem cells (hMSCs) toward myogenic phenotypes in a cardiac niche, the present study aimed to determine the role of miR-16 in this process.Main methods: hMSCs and neonatal rat ventricular myocytes were co-cultured indirectly in two chambers to set up a cardiac microenvironment (niche). miRNA expression profile in cardiac-niche-induced hMSCs was detected by miRNA microarray. Cardiac marker expression and cell cycle analysis were determined in different treatment hMSCs. Quantitative real-time PCR and Western blot were used to identify the expression of mRNA, mature miRNA and protein of interest.Key findings: miRNA dysregulation was shown in hMSCs after cardiac niche induction. miR-16 was upregulated in cardiac-niche-induced hMSCs. Overexpression of miR-16 significantly increased G1-phase arrest of the cell cycle in hMSCs and enhanced the expression of cardiac marker genes, including GATA4, NK2-5, MEF2C and TNNI3. Differentiation-inducing factor 3 (DIF-3), a G0/G1 cell cycle arrest compound, was used to induce G1 phase arrest in cardiac-niche-induced hMSCs, and the expression of cardiac marker genes was upregulated in DIF-3-treated hMSCs. The expression of CCND1, CCND2 and CDK6 was suppressed by miR-16 in hMSCs. CDK6, CCND1 or CCND2 knockdown resulted in G1 phase arrest in hMSCs and upregulation of cardiac marker gene expression in hMSCs in a cardiac niche.Significance: miR-16 enhances G1 phase arrest in hMSCs, contributing to the differentiation of hMSCs toward myogenic phenotypes when in a cardiac niche. This mechanism provides a novel strategy for pre-modification of hMSCs before hMSC-based transplantation therapy for severe heart diseases. Crown Copyright (c) 2012 Published by Elsevier Inc. All rights reserved.