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MicroRNA-320 regulates matrix metalloproteinase-13 expression in chondrogenesis and interleukin-1β-induced

F Meng¹, Z Zhang¹, W Chen¹

¹Department of Joint Surgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510080, China.

Osteoarthritis and Cartilage

|January 18, 2016

Abstract

Objective:

Metalloproteinases (MMPs) are key regulators of osteoarthritis (OA) and collagen degradation and have been shown to participate in endochondral ossification. The aim of this study was to determine whether microRNA-320 (miR-320) regulates the expression of MMP-13 in chondrogenesis and inflammation.

Experimental Design:

miR-320 expression was assessed in vitro, in the ATDC5 cell model of chondrogenesis and in interleukin-1β (IL-1β)-treated primary mouse chondrocytes (PMCs), and in vivo, in normal and OA human cartilage by in situ hybridization. ATDC5 and PMCs were transfected with miR-320 or its antisense inhibitor (anti-miR-320), respectively. The roles of activated MAP kinases (MAPK) and NF-κB were evaluated by using specific inhibitors. Direct interaction between miR-320 and its putative binding site in the 3'-untranslated region (3'-UTR) of Mmp-13 mRNA was confirmed by the luciferase reporter assay.

Results:

miR-320 expression was elevated in chondrogenic and hypertrophic ATDC5, while significantly reduced in OA cartilage compared with normal cartilage. Stimulation with IL-1β led to a significant reduction in miR-320 expression in PMCs. Upregulation of MMP-13 expression was correlated with downregulation of miR-320 expression in both PMCs and ATDC5. Overexpression of miR-320 suppressed the activity of a reporter construct containing the 3'-UTR and inhibited MMP-13 expression in both ATDC5 and IL-1β-treated PMCs, while treatment with anti-miR-320 enhanced MMP-13 expression. NF-κB and MAPK activation downregulated miR-320 expression.

Conclusion:

Cartilage development and homeostasis are influenced by miR-320, which directly targets MMP-13 and regulates chondrogenesis and the IL-1β-stimulated catabolic effect in mouse chondrocytes.

Insights

MicroRNA-320 (miR-320) regulates matrix metalloproteinase-13 (MMP-13) in cartilage development and osteoarthritis. Upregulating miR-320 suppresses MMP-13, impacting chondrogenesis and inflammation.

Area of Science:

Molecular Biology
Biochemistry
Cell Biology

Background:

Matrix metalloproteinases (MMPs), particularly MMP-13, are crucial in collagen degradation and osteoarthritis (OA) pathogenesis.
MMPs also play a role in endochondral ossification, a process relevant to cartilage development and homeostasis.

Purpose of the Study:

To investigate the regulatory role of microRNA-320 (miR-320) in chondrogenesis and inflammation.
To determine if miR-320 directly influences the expression of MMP-13.

Main Methods:

Assessed miR-320 expression in chondrogenic (ATDC5) and primary mouse chondrocytes (PMCs) in vitro, and in human cartilage in vivo.
Utilized transfection with miR-320 or anti-miR-320 in ATDC5 and PMCs.
Investigated the involvement of MAP kinase (MAPK) and NF-κB signaling pathways.
Confirmed direct interaction between miR-320 and MMP-13 mRNA using a luciferase reporter assay.

Main Results:

miR-320 expression was elevated in chondrogenic cells but reduced in OA cartilage.
Interleukin-1β (IL-1β) stimulation decreased miR-320 expression in PMCs.
miR-320 overexpression suppressed MMP-13 expression, while inhibition of miR-320 increased it.
NF-κB and MAPK activation led to decreased miR-320 expression.

Conclusions:

miR-320 directly targets MMP-13 mRNA, regulating its expression.
miR-320 influences cartilage development and homeostasis.
miR-320 plays a role in mitigating the catabolic effects of IL-1β in chondrocytes.

Read full analysis

Keywords:

Chondrocytes Chondrogenesis Metalloproteinase-13 MiRNA-320 Osteoarthritis

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