scholarly journals CCAAT/ENHANCER binding protein β mediates expression of matrix metalloproteinase 13 in human articular chondrocytes in inflammatory arthritis

2009 ◽  
Vol 60 (3) ◽  
pp. 708-716 ◽  
Author(s):  
Mitsumasa Hayashida ◽  
Ken Okazaki ◽  
Junichi Fukushi ◽  
Akio Sakamoto ◽  
Yukihide Iwamoto
Keyword(s):  
Matrix Metalloproteinase ◽  
Inflammatory Arthritis ◽  
Articular Chondrocytes ◽  
Binding Protein ◽  
Human Articular Chondrocytes ◽  
Matrix Metalloproteinase 13 ◽  
Enhancer Binding Protein ◽  
Ccaat Enhancer Binding Protein

scholarly journals CCAAT/enhancer binding protein β regulates expression of matrix metalloproteinase-3 in arthritis

2011 ◽  
Vol 71 (1) ◽  
pp. 99-107 ◽  
Author(s):  
Hidetoshi Tsushima ◽  
Ken Okazaki ◽  
Mitsumasa Hayashida ◽  
Takahiro Ushijima ◽  
Yukihide Iwamoto
Keyword(s):  
Matrix Metalloproteinase ◽  
Chromatin Immunoprecipitation ◽  
Binding Protein ◽  
Chromatin Immunoprecipitation Assay ◽  
Immunoprecipitation Assay ◽  
Matrix Metalloproteinase 3 ◽  
Enhancer Binding Protein ◽  
Ccaat Enhancer Binding Protein ◽  
And Cartilage ◽  
In Cells

ObjectivesTo investigate whether CCAAT/enhancer binding protein β (C/EBPβ) mediates the expression of matrix metalloproteinase-3 (MMP-3) and aggrecanases in arthritis.MethodsLocalisation of C/EBPβ and MMP-3 in synovium and cartilage from patients with rheumatoid arthritis and osteoarthritis was determined by immunohistochemistry. Cell lines SW982, C28/I2 and human fibroblast-like synoviocytes stimulated by interleukin 1β (IL-1β) were subjected to western blotting and quantitative PCR. Overexpression of C/EBPβ by adenovirus was performed in cells and organ culture of normal cartilage. Knockdown of C/EBPβ by small interference RNA was performed in cells. Activity of the human MMP-3 and aggrecanase-2 ADAMTS-5 (a disintegrin and metalloproteinase with thrombospondin motifs) promoters was analysed by a luciferase assay. To determine whether C/EBPβ directly binds to the MMP-3 or ADAMTS-5 promoter,a chromatin immunoprecipitation assay was performed.ResultsImmunohistochemistry showed that C/EBPβ and MMP-3 were co-localised in arthritic synovium and cartilage. Western blots revealed increased C/EBPβ expression in cells treated with IL-1β. Expression of MMP-3, MMP-13 and ADAMTS-5 mRNA was significantly increased by the overexpression of C/EBPβ. C/EBPβ stimulated MMP-3 expression and induced matrix degradation in cartilage explants. C/EBPβ knockdown reduced MMP-3 and ADAMTS-5 expression. C/EBPβ stimulated the 2011 bp MMP-3 promoter and the 1768 bp ADAMTS-5 promoter in a dose-dependent manner. Deletion and mutation analysis of the MMP-3 promoter showed that the C/EBPβ core responsive element was located between −108 bp and −100 bp. The chromatin immunoprecipitation assay showed that C/EBPβ was directly bound to MMP-3 and ADAMTS-5 promoters.ConclusionsThese data demonstrate that C/EBPβ is involved in expression of MMP-3 and ADAMTS-5 in arthritic synovium and cartilage.

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