Presence and function of microRNA-92a in chondrogenic ATDC5 and adipose-derived mesenchymal stem cells.
Presence and function of microRNA-92a in chondrogenic ATDC5 and adipose-derived mesenchymal stem cells.
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软骨形成 ATDC5 和脂肪间充质干细胞中 microRNA-92a 的存在和功能
DOI：
10.3892/mmr.2015.4008
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发表时间：
2015-10
影响因子：
3.4
通讯作者：
Liao W
中科院分区：
文献类型：
--
作者：
Hou C;Zhang Z;Zhang Z;Wu P;Zhao X;Fu M;Sheng P;Kang Y;Liao W
本研究旨在探讨microRNA-92 a（miR-92 a）在软骨发生和软骨退变中的存在及其生物学功能。将人脂肪间充质干细胞（hADSCs）诱导成软骨，并将原代人/小鼠软骨细胞（PHCs/PMCs）和成软骨ATDC 5细胞用白细胞介素-1 β（IL-1β）刺激。使用lipofectamine 2000将miR-92 a模拟物/抑制剂转染到ATDC 5细胞中。采用逆转录-定量聚合酶链反应分析基因表达。用阿辛蓝染色观察软骨结节和软骨样微团。利用米兰达、miRDB、CLIP-Seq、TargetScan和京都基因与基因组百科全书等技术对miR-92 a的潜在靶基因、信号通路和功能进行了研究。miR-92 a在软骨形成ATDC 5细胞和hADSC中的表达升高，并且在IL-1β诱导的ATDC 5细胞、PMC和PHCs中也升高。miR-92 a的强制表达增强了col 9a 2和聚集蛋白聚糖的表达水平。共预测了279个基因为miR-92 a的潜在靶基因。磷酸肌醇3-激酶（PI 3 K）-Akt、ErbB和黏着斑激酶通路、细胞外基质（ECM）-受体相互作用和哺乳动物雷帕霉素靶蛋白（mTOR）信号通路被认为介导miR-92 a对软骨形成和软骨退变的作用。这些结果表明，miR-92 a参与软骨形成和IL-1β诱导的软骨细胞反应。miR-92 a促进col 9a 2和聚集蛋白聚糖的表达。
The aim of the present study was to investigate the presence and biological function of microRNA-92a (miR-92a) in chondrogenesis and cartilage degeneration. Human adipose-derived mesenchymal stem cells (hADSCs) in micromass and chondrocyte-like ATDC5 cells were induced to chondrogenesis, and primary human/mouse chondrocytes (PHCs/PMCs) and chondrogenic ATDC5 cells were stimulated with interleukin-1β (IL-1β). An miR-92a mimic/inhibitor was transfected into the ATDC5 cells using lipofectamine 2000. Gene expression was analyzed using reverse transcription-quantitative polymerase chain reaction. Alcian blue was used to stain the cartilage nodules and chondrogenic micromass. The potential target genes, signaling pathways and functions of miR-92a were examined using miRanda, miRDB, CLIP-Seq, TargetScan and Kyoto Encyclopedia of Genes and Genomes. The expression of miR-92a was elevated in the chondrogenic ATDC5 cells and hADSCs, and also in the IL-1β-induced ATDC5 cells, PMCs and PHCs. Forced expression of miR-92a enhanced the expression levels of col9a2 and aggrecan. A total of 279 genes were predicted as potential target genes of miR-92a. The phosphoinositide 3-kinase/PI3K)-Akt, ErbB and focal adhesion kinase pathways, extracellular matrix (ECM)-receptor interaction and the mammalian target of rapamycin (mTOR) signaling pathway were suggested to mediate the effects of miR-92a on chondrogenesis and cartilage degeneration. These results demonstrated that miR-92a was involved in chondrogenesis and the chondrocyte response induced by IL-1β. miR-92a positively contributed to the expression of col9a2 and of aggrecan.