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长链非编码RNA CIR通过作为Mir-27b的海绵促进骨关节炎中软骨细胞外基质降解。

Long Noncoding RNA CIR Promotes Chondrocyte Extracellular Matrix Degradation in Osteoarthritis by Acting as a Sponge For Mir-27b.

作者信息

Li Yu-Fei, Li Shu-Hua, Liu Yong, Luo Ya-Tong

出版信息

Cell Physiol Biochem. 2017;43(2):602-610. doi: 10.1159/000480532. Epub 2017 Sep 21.

Abstract

BACKGROUND/AIMS: Osteoarthritis (OA) is a degenerative joint disease characterized by articular cartilage degradation. The degradation of the extracellular matrix (ECM) of chondrocyte is closely associated with the destruction of joints in OA patients. lncRNAs are non-coding segments of RNA that possess important regulatory functions at the cellular level and in a variety of pathophysiological processes. The present study was conducted to investigate whether lncRNA-CIR regulated the expression of MMP13 as a sponge of miR-27 in OA.

METHODS

Primary cultured chondrocytes were challenged by IL-1β and TNF-α to simulate OA conditions. qRT-PCR was performed to detect the miR-27, lncRNA-CIR, MMP13 mRNA expression levels. Western blot was applied to detect MMP13 protein expression. Soluble sGAG secretion/ formation was analysed by the dimethylmethylene blue (DMMB) assay. lncRNA-CIR overexpression or inhibition was performed using overexpression plasmid and small interfering RNAs (siRNAs), respectively.

RESULTS

lncRNA-CIR significantly up-regulated in OA patients, concomitantly down-regulated miR-27 and up-regulated MMP13. Bioinformatics analysis predicted miR-27 was the target of both lncRNA-CIR and MMP13. Overexpression of lncRNA-CIR significantly increased the expression of MMP13, while miR-27 remarkably suppressed the expression of MMP13, Accompanying with the increases of mRNA level, protein level and relative luciferase activity.

CONCLUSION

The present findings indicated that lncRNA-CIR/miR-27/MMP13 axis involved in the degradation of the ECM of chondrocyte in OA.

摘要

背景/目的:骨关节炎(OA)是一种以关节软骨退变 为特征的退行性关节疾病。软骨细胞外基质(ECM)的退变与OA患者关节破坏密切相关。长链非编码RNA(lncRNA)是RNA的非编码片段,在细胞水平和多种病理生理过程中具有重要的调控功能。本研究旨在探讨lncRNA-CIR在OA中是否作为miR-27的海绵调节MMP13的表达。

方法

用白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)刺激原代培养的软骨细胞以模拟OA状态。采用qRT-PCR检测miR-27、lncRNA-CIR、MMP13 mRNA表达水平。应用蛋白质免疫印迹法检测MMP13蛋白表达。采用二甲基亚甲基蓝(DMMB)法分析可溶性糖胺聚糖(sGAG)的分泌/形成。分别使用过表达质粒和小干扰RNA(siRNA)进行lncRNA-CIR的过表达或抑制。

结果

OA患者lncRNA-CIR显著上调,同时miR-27下调,MMP13上调。生物信息学分析预测miR-27是lncRNA-CIR和MMP13的靶点。lncRNA-CIR过表达显著增加MMP13的表达,而miR-27显著抑制MMP13的表达,同时伴随着mRNA水平、蛋白水平和相对荧光素酶活性的增加。

结论

本研究结果表明lncRNA-CIR/miR-27/MMP13轴参与OA中软骨细胞ECM的降解。

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