MiRNA-140 is a negative feedback regulator of MMP-13 in IL-1ß-stimulated human articular chondrocyte C28/I2 cells.

ABSTRACT

OBJECTIVE: Osteoarthritis is a degenerative joint disease, in which matrix metalloproteinase (MMP)-13 plays an important role. This study aimed to investigate miRNA-140-mediated negative regulation of MMP-13 expression in interleukin-1ß (IL-1ß)-stimulated human cartilage cells. METHODS: The human cartilage cell line C28/I2 was cultured in the presence of IL-1ß to mimic an osteoarthritic environment. Expression of miRNA-140 and MMP-13 was analyzed after 48 h by real-time RT-PCR and western blot analyses. MiRNA-140 mediated regulation of MMP-13 expression was analyzed by luciferase reporter assays and anti-miRNA-140 oligonucleotide transfection. Furthermore, miRNA-140 and MMP-13 expression was analyzed following DHMEQ treatment. RESULTS: Expression of miRNA-140 and MMP-13 was elevated in IL-1ß-stimulated C28/I2 cells. Bioinformatic prediction showed that the 3'-UTR of MMP-13 mRNA contained a potential binding miRNA-140 site and luciferase mRNA fused with 3'-UTR of MMP-13 mRNA was shown to be repressed by miRNA-140 in reporter assays. Expression of MMP-13 was elevated in IL-1ß-stimulated C28/I2 cells following anti-miRNA-140 oligonucleotide transfection. NF-κB activity was inhibited in DHMEQ treated IL-1ß-stimulated C28/I2 cells and was associated with decreased miRNA-140 and MMP-13 expression. CONCLUSION: Expression of miRNA-140 and MMP-13 was induced by IL-1ß. Expression of miRNA-140 inhibited MMP-13 in C28/I2 cells. Expression of miRNA-140 and MMP-13 was shown to be NF-κB-dependent.