title: MiRNA-140 is a negative feedback regulator of MMP-13 in IL-1β-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-1β (IL-1β)-stimulated human cartilage cells.
  
  
  METHODS
  The human cartilage cell line C28/I2 was cultured in the presence of IL-1β 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-1β-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-1β-stimulated C28/I2 cells following anti-miRNA-140 oligonucleotide transfection. NF-κB activity was inhibited in DHMEQ treated IL-1β-stimulated C28/I2 cells and was associated with decreased miRNA-140 and MMP-13 expression.
  
  
  CONCLUSIONS
  Expression of miRNA-140 and MMP-13 was induced by IL-1β. 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.
authors.affiliation: Department of Orthopaedics, The 3rd Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China.
authors.forename: Zu-Jian
authors.initials: ZJ
authors.lastname: Liang
authors.forename: Hong
authors.initials: H
authors.lastname: Zhuang
authors.forename: Guo-Xia
authors.initials: GX
authors.lastname: Wang
authors.forename: Zhao
authors.initials: Z
authors.lastname: Li
authors.forename: Huan-Tian
authors.initials: HT
authors.lastname: Zhang
authors.forename: Tian-Qi
authors.initials: TQ
authors.lastname: Yu
authors.forename: Bai-Dang
authors.initials: BD
authors.lastname: Zhang
mesh_terms: D002356:Cartilage / Q000201:enzymology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000201:enzymology*; D025461:Feedback, Physiological; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology*; D053509:Matrix Metalloproteinase 13 / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D035683:MicroRNAs / Q000032:analysis / Q000502:physiology*; D016328:NF-kappa B / Q000502:physiology; D010003:Osteoarthritis / Q000201:enzymology* / Q000209:etiology
chemical_list: D053583:Interleukin-1beta; D035683:MicroRNAs; C544302:Mirn140 microRNA, human; D016328:NF-kappa B; D053509:Matrix Metalloproteinase 13