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Arthritis Rheum. 2009 Mar;60(3):708-16. doi: 10.1002/art.24332.

CCAAT/enhancer binding protein beta mediates expression of matrix metalloproteinase 13 in human articular chondrocytes in inflammatory arthritis.

Author information

1
Kyushu University, Fukuoka, Japan.

Abstract

OBJECTIVE:

To determine the function of CCAAT/enhancer binding protein beta (C/EBPbeta) in the expression of matrix metalloproteinase 13 (MMP-13) in chondrocytes in inflammatory arthritis.

METHODS:

Cartilage obtained from patients with rheumatoid arthritis and osteoarthritis was immunostained for expression of C/EBPbeta or MMP-13. Interleukin-1beta- or tumor necrosis factor alpha (TNFalpha)-stimulated chondrocytes were subjected to Western blotting and real-time reverse transcriptase-polymerase chain reaction (RT-PCR). MMP-13 promoter assays were conducted, and the C/EBPbeta response element was characterized by deletion and mutation analysis. C-28/I2 cells were treated with TNFalpha and subjected to chromatin immunoprecipitation (ChIP) assays. Finally, C/EBPbeta-liver-enriched activator protein (LAP) was overexpressed in C-28/I2 cells or cartilage tissues, and MMP-13 expression was analyzed.

RESULTS:

C/EBPbeta and MMP-13 expression was colocalized in chondrocytes in arthritic cartilage. MMP-13 promoter activity was stimulated by C/EBPbeta overexpression in a dose-dependent manner. Luciferase assays revealed that a -981-bp promoter had the greatest activity, while deletion to -936 bp strongly diminished promoter activity. Luciferase activity was repressed to basal levels by mutations in potential C/EBP binding sites. The stimulatory effects of C/EBPbeta overexpression were diminished by mutation. ChIP assays revealed that TNFalpha treatment enhanced the binding of C/EBPbeta to the MMP-13 promoter. When C/EBPbeta-LAP was overexpressed in C-28/I2 cells, endogenous MMP-13 expression was stimulated up to 32-fold as detected by real-time RT-PCR. Furthermore, following adenoviral overexpression of C/EBPbeta-LAP in organ culture of articular cartilage, stimulation of MMP-13 was also detected by immunohistochemistry.

CONCLUSION:

C/EBPbeta directly binds to the MMP-13 promoter region and stimulates the expression of MMP-13 in chondrocytes in inflammatory arthritis.

PMID:
19248099
DOI:
10.1002/art.24332
[Indexed for MEDLINE]
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