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

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.

The Protective Effect of Ligustilide in Osteoarthritis: An in Vitro and in Vivo Study

“…Inhibition of the degradation of collagen II and aggrecan may have positive effects on OA treatment. Type II collagen is cleaved by MMPs, whereas aggrecan is cleaved by ADAMTS [39][40][41]. In our study, in the presence of IL-β, LIG not only promoted the synthesis of type II collagen and aggrecan, but also inhibited the catabolism of ECM components by downregulating MMPs and ADAMTS-5, which helped to maintain ECM homeostasis.…”

Onset and Progression of Human Osteoarthritis—Can Growth Factors, Inflammatory Cytokines, or Differential miRNA Expression Concomitantly Induce Proliferation, ECM Degradation, and Inflammation in Articular Cartilage?

“…MiR-27b, which directly targets MMP-13 , is downregulated in human knee OA AC, compared to AC from young and healthy AC donors [167]. Hydrostatic pressure increases both miR-27a and miR-27b expression specifically in human hip OA AC monolayer cell cultures, but not in cell cultures derived from normal hip AC [168].…”

“…Remarkably, though rIL-1β downregulates miR-27a and miR-27b in human late stage knee OA AC explant cultures, in synovial explant cultures of patients with late stage knee OA AC an upregulation and enhanced secretion of miR-27a and miR-27b was detectable upon rIL-1β stimulation [159]. Long non-coding RNA-cartilage injury-related (lncRNA-CIR), which is upregulated in OA AC, acts as a sponge for miR-27, whereas miR-27 directly represses lncRNA-CIR expression [167]. In addition, in human chondrosarcoma cells, expression of miR-27b is negatively regulated by adiponectin (ADPN), an adipokine [169].…”

miR-27b Suppresses Endothelial Cell Proliferation and Migration by Targeting Smad7 in Kawasaki Disease

“…Smad7 mRNA fragment (from 771 to 830), which was located in CDS of Smad7, was amplified and cloned into the luciferase repoter via SpeI and HindIII sites. Briefly, cells plated in a 48-well plate were co-transfected with 50 nM miRNA mimics or negative control oligonucleotides (NC), 50 ng of firefly luciferase reporter and 10 ng of pRL-TK (Promega, USA) using the JetPRIME reagent (Polyplus-transfection) [10]. Cells were collected 36 hours after transfection and analyzed using DualLuciferase Reporter Assay System (Promega, USA).…”