MicroRNA‐27b regulates the expression of matrix metalloproteinase 13 in human osteoarthritis chondrocytes

Abstract

Objective Aberrant posttranscriptional regulation of matrix metalloproteinases (MMPs) by microRNA has emerged as an important factor in human diseases. The aim of this study was to determine whether the expression of MMP-13 in human osteoarthritis (OA) chondrocytes is regulated by microRNA. Methods Chondrocytes were stimulated with interleukin-1β (IL-1β) in vitro. Total RNA was prepared using TRIzol reagent. Polymerase chain reaction (PCR)–based arrays were used to determine the expression profile of 352 human microRNA. Gene expression was quantified using TaqMan assays, and microRNA targets were identified using bioinformatics. Transfection with reporter construct and microRNA mimic was used to verify suppression of target messenger RNA (mRNA). Gene expression of argonaute and Dicer was determined by reverse transcription–PCR, and expression of protein was determined by immunoblotting. The role of activated MAP kinases (MAPKs) and NF-κB was evaluated using specific inhibitors. Results In IL-1β–stimulated OA chondrocytes, 42 microRNA were down-regulated, 2 microRNA were up-regulated, and the expression of 308 microRNA remained unchanged. In silico analysis identified a sequence in the 3′-untranslated region (3′-UTR) of MMP-13 mRNA complementary to the seed sequence of microRNA-27b (miR-27b). Increased expression of MMP-13 correlated with down-regulation of miR-27b. Overexpression of miR-27b suppressed the activity of a reporter construct containing the 3′-UTR of human MMP-13 mRNA and inhibited the IL-1β–induced expression of MMP-13 protein in chondrocytes. NF-κB and MAPK activation down-regulated the expression of miR-27b. Conclusion Our data demonstrated the expression of miR-27b in both normal and OA chondrocytes. Furthermore, IL-1β–induced activation of signal transduction pathways associated with the expression of MMP-13 down-regulated the expression of miR-27b. Thus, miR-27b may play a role in regulating the expression of MMP-13 in human chondrocytes.

Human matrix metalloproteinases: An ubiquitarian class of enzymes involved in several pathological processes

“…The chondroprotective effects of micro-RNA inhibition of the MMP-13 expression was proved in human cartilage explants in vitro and through the development of inflammatory arthritis in mouse model in vivo (Akhtar et al, 2010).…”

MicroRNAs and post-transcriptional regulation of skeletal development

“…At present, therapy is usually based on symptomatic treatment, mainly using nonsteroidal anti-inflammatory drugs (NSAIDs), but these fail to slow articular degeneration and disease progression. Studies on OA have demonstrated that miRNA expression is regulated during this process, indicating the possibility of future miRNA therapies (Iliopoulos et al 2008, Jones et al 2009, Akhtar et al 2010, Diaz-Prado et al 2012. One of the main characteristics of osteoarthritic chondrocytes is the secretion of the pro-inflammatory cytokines interleukin 1b (IL1b) and TNFa.…”

“…Numerous miRNAs have been shown to be regulated by IL1b action (Miyaki et al 2009, Dai et al 2012, Matsukawa et al 2013. MMP13 is regulated directly by miR-27b and miR-127-5p (Akhtar et al 2010 or indirectly by miR-27a, miR-9, miR-488, or miR-22 (Iliopoulos et al 2008, Jones et al 2009, Tardif et al 2009, Song et al 2013b, among others.…”

Next-generation Sequencing Identifies Articular Cartilage and Subchondral Bone Mirnas after ESWT on Early Osteoarthritis Knee

“…Several miRNAs including miR-140, miR-199a, mir-193 and mir-29a/29b control the anabolic and catabolic regulation in cartilage [42]. In human OA chondrocytes, miR-27b controls the expression of matrix metalloproteinase 13(MMP13) in OA treatment [43]. MiR-181a is reported to control osteopontin expression in cancer cell and inhibit expression in bone marrow-derived mesenchymal stem cells to maintain bone remodeling balance [44,45].…”