MicroRNA-92a-3p regulates the expression of cartilage-specific genes by directly targeting histone deacetylase 2 in chondrogenesis and degradation
MicroRNA-92a-3p regulates the expression of cartilage-specific genes by directly targeting histone deacetylase 2 in chondrogenesis and degradation
复制标题
MicroRNA-92a-3p 通过在软骨形成和降解过程中直接靶向组蛋白脱乙酰酶 2 来调节软骨特异性基因的表达。
DOI：
10.1016/j.joca.2016.11.006
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发表时间：
2017-04-01
影响因子：
7
通讯作者：
Kang, Y.
中科院分区：
文献类型：
--
作者：
Mao, G.;Zhang, Z.;Kang, Y.
目的：在骨关节炎（OA）和软骨基质降解患者中发现组蛋白脱乙酰酶2（HDAC 2）活性增加，并已显示其介导人软骨细胞中软骨特异性基因表达的抑制。我们的目的是确定是否microRNA-92 a-3 p（miR-92 a-3 p）调节软骨特异性基因的表达，通过有针对性的HDAC 2在chondrogenesis和degradation.Methods：miR-92 a-3 p的表达进行了评估，在体外的人骨髓间充质干细胞（hMSCs）模型的软骨形成和正常和OA原代人软骨细胞（PHCs），并在正常和OA人软骨原位杂交。分别用miR-92 a-3 p或其反义抑制剂（anti-miR-92 a-3 p）转染hMSC和PHCs。将PHCs用miR-92 a-3 p或抗miR-92 a-3 p转染24小时，然后用抗ac-H3抗体进行染色质免疫沉淀（ChIP）测定。结果：miR-92 a-3 p在成软骨和肥大hMSC中的表达明显增加，而在OA软骨中的表达明显减少。miR-92 a-3 p的过表达抑制了含有3 '-UTR的报告构建体的活性，并抑制了hMSC和PHCs中的HDAC 2表达，而用抗miR-92 a-3 p处理增强了HDAC 2表达。结论：miR-92 a-3 p调控软骨发育和稳态，其作用靶点为HDAC 2，提示组蛋白乙酰化在软骨基质表达增加中起重要作用。(C)2016国际骨关节炎研究学会。由爱思唯尔有限公司出版。保留所有权利。
Objective: Increased activity of histone deacetylase 2 (HDAC2) has been found in patients with osteoarthritis (OA) and cartilage matrix degradation and has been shown to mediate the repression of cartilage-specific gene expression in human chondrocytes. We aimed to determine whether microRNA-92a-3p (miR-92a-3p) regulates cartilage-specific gene expression via targeted HDAC2 in chondrogenesis and degradation.Methods: miR-92a-3p expression was assessed in vitro in a human mesenchymal stem cells (hMSCs) model of chondrogenesis and in normal and OA primary human chondrocytes (PHCs), and in normal and OA human cartilage by in situ hybridization. hMSCs and PHCs were transfected with miR-92a-3p or its antisense inhibitor (anti-miR-92a-3p), respectively. PHCs were transfected with miR-92a-3p or anti-miR-92a-3p for 24 h before chromatin immunoprecipitation (ChIP) assay was performed with anti-ac-H3 antibody. Direct interaction between miR-92a-3p and its putative binding site in the 3'-untranslated region (3'-UTR) of HDAC2 mRNA was confirmed by luciferase reporter assay.Results: miR-92a-3p expression was elevated in chondrogenic and hypertrophic hMSC, while reduced in OA cartilage compared with normal cartilage. The overexpression of miR-92a-3p suppressed the activity of a reporter construct containing the 3'-UTR and inhibited HDAC2 expression in both hMSCs and PHCs, while treatment with anti-miR-92a-3p enhanced HDAC2 expression. ChIP assays showed that miR-92a-3p enhances H3 acetylation on aggrecan (ACAN), cartilage oligomeric protein (COMP) and Col2a1 promoter, and also promotes relative cartilage matrix expression.Conclusion: Our results suggest that miR-92a-3p regulates cartilage development and homeostasis, which directly targets HDAC2, indicating histone hyperacetylation plays an important role in increased expression of cartilage matrix. (C) 2016 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.