title: MicroRNA-127-5p regulates matrix metalloproteinase 13 expression and interleukin-1β-induced catabolic effects in human chondrocytes.
abstract: OBJECTIVE
  MicroRNAs (miRNAs), small noncoding RNA molecules, are involved in the pathogenesis of various diseases such as cancer and arthritis. The aim of this study was to determine whether miR-127-5p regulates interleukin-1β (IL-1β)-induced expression of matrix metalloproteinase 13 (MMP-13) and other catabolic factors in human chondrocytes.
  
  
  METHODS
  Expression of miR-127-5p and MMP-13 by normal and osteoarthritic (OA) human cartilage was determined using real-time polymerase chain reaction. The effect of miR-127-5p on MMP-13 expression was evaluated using transient transfection of human chondrocytes or chondrogenic SW-1353 cells with miR-127-5p or its antisense inhibitor (anti-miR-127-5p). MMP-13 protein production was quantified by enzyme-linked immunosorbent assay, and the involvement of miR-127-5p in IL-1β-mediated catabolic effects was examined by immunoblotting. MicroRNA-127-5p binding with the putative site in the 3'-untranslated region (3'-UTR) of MMP-13 messenger RNA (mRNA) was validated by luciferase reporter assay.
  
  
  RESULTS
  There was a significant reduction in miR-127-5p expression in OA cartilage compared with normal cartilage. Up-regulation of MMP-13 expression by IL-1β was correlated with down-regulation of miR-127-5p expression in human chondrocytes. MicroRNA-127-5p suppressed IL-1β-induced MMP-13 production as well as the activity of a reporter construct containing the 3'-UTR of human MMP-13 mRNA. In addition, mutation of the miR-127-5p binding site in the 3'-UTR of MMP-13 mRNA abolished miR-127-5p-mediated repression of reporter activity. Conversely, treatment with anti-miR-127-5p remarkably increased reporter activity and MMP-13 production. Interestingly, the IL-1β-induced activation of JNK, p38, and NF-κB and expression of MMP-1 and cyclooxygenase 2 were significantly inhibited by miR-127-5p.
  
  
  CONCLUSIONS
  MicroRNA-127-5p is an important regulator of MMP-13 in human chondrocytes and may contribute to the development of OA.
authors.affiliation: Hallym University Sacred Heart Hospital, Kyunggi, Republic of Korea.
authors.forename: Su Jin
authors.initials: SJ
authors.lastname: Park
authors.forename: Eun Jeong
authors.initials: EJ
authors.lastname: Cheon
authors.forename: Mi Hyun
authors.initials: MH
authors.lastname: Lee
authors.forename: Hyun Ah
authors.initials: HA
authors.lastname: Kim
mesh_terms: D002358:Cartilage, Articular / Q000187:drug effects / Q000378:metabolism*; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D053583:Interleukin-1beta / Q000494:pharmacology*; D053509:Matrix Metalloproteinase 13 / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D010003:Osteoarthritis / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology
chemical_list: D053583:Interleukin-1beta; C547276:MIRN127 microRNA, human; D035683:MicroRNAs; D016328:NF-kappa B; D053509:Matrix Metalloproteinase 13