Role of miR-92a in chondrogenic ATDC5 and adipose derived mesenchymal stem cells, and in chondrocytes with interleukin-1beta
Role of miR-92a in chondrogenic ATDC5 and adipose derived mesenchymal stem cells, and in chondrocytes with interleukin-1beta
复制标题
miR-92a 在软骨形成 ATDC5 和脂肪源性间充质干细胞以及具有 IL-1β 的软骨细胞中的作用
作者：
Hou C;Zhang Z;Zhang Z;Wu P;Zhao X;Fu M;Kang Y;Liao W
摘要目的探讨miR-92 a在软骨形成和软骨退变中的表达及其生物学功能。方法分别用TGF-β和ITS+ Premix诱导微团脂肪间充质干细胞（hADSCs）和软骨样ATDC 5细胞向软骨分化。原代人/小鼠软骨细胞（PHCs/PMCs）和软骨形成ATDC 5用白细胞介素-1 β（IL-1b）刺激。用lipofectamine 2000将miR-92 a模拟物/抑制剂转染到ATDC 5细胞中。用qPCR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和京都基因和基因组百科全书（KEGG）预测miR-92 a的潜在靶基因、信号通路和功能。结果MiR-92 a在成软骨的ATDC 5和hADSC中以及在IL-1b诱导的ATDC 5、PMC和PHCs中表达增加。miR-92 a的强制表达增强了col 9a 2和聚集蛋白聚糖的表达。共预测了279个基因为miR-92 a的潜在靶基因。预测PI 3 K-Akt、ErbB、粘着斑激酶通路、ECM-受体相互作用和mTOR信号通路介导miR-92 a对软骨形成和软骨退变的作用。结论发现miR-92 a在IL-1b诱导的软骨形成和软骨细胞反应中发挥作用。miR-92 a促进col 9a 2和聚集蛋白聚糖的表达。预计PI 3 K-Akt、ErbB、粘着斑激酶途径、ECM-受体相互作用和mTOR信号途径可能介导miR-92 a对软骨形成和软骨变性的影响。
Abstract Purpose To investigate the expression and biological function of miR-92a in chondrogenesis and cartilage degeneration. Methods Micromass of adipose derived mesenchymal stem cells (hADSCs) and chondrocyte-like ATDC5 cell line were induced to chondrogenesis with TGF-beta and ITS+ Premix respectively. Primary human/mouse chondrocytes (PHCs/PMCs) and chondrogenic ATDC5 were stimulated with interleukin-1beta (IL-1b). MiR-92a mimic/inhibitor was transfected into ATDC5 cells with lipofectamine 2000. Gene expression was analyzed with qPCR. Alcian blue were used to stain the cartilage nodules and chondrogenic micromass. Potential target genes, signaling pathways, and functions of miR-92a were predicted with miRanda, miRDB, CLIP-Seq, TargetScan, and Kyoto Encyclopedia of Genes and Genomes (KEGG). Results MiR-92a expression was elevated in chondrogenic ATDC5 and hADSCs, as well as in IL-1b induced ATDC5, PMCs and PHCs. Forced expression of miR-92a enhanced expression of col9a2 and aggrecan. A total of 279 genes was predicted as potential target genes of miR-92a. PI3K-Akt, ErbB, Focal adhesion kinase pathway, ECM-receptor interaction, and mTOR signaling pathway were predicted to mediate the effect of miR-92a on chondrogenesis and cartilage degeneration. Conclusions We found the role of miR-92a in chondrogenesis and chondrocyte response induced with IL-1b. MiR-92a positively contributed to col9a2 and aggrecan expression. PI3K-Akt, ErbB, Focal adhesion kinase pathway, ECM-receptor interaction, and mTOR signaling pathway were possibly predicted to mediate the effect of miR-92a on chondrogenesis and cartilage degeneration.