RelA is required for IL-1beta stimulation of Matrix Metalloproteinase-1 expression in chondrocytes.
OBJECTIVE
Interleukin-1beta (IL-1beta) stimulates collagenase-1 (Matrix Metalloproteinase-1 (MMP-1)) expression in articular chondrocytes, leading to cleavage of type II collagen and irreversible cartilage degradation. The nuclear factor-kappa B (NF-kappaB) pathway is potently activated in IL-1beta-stimulated cells and has been implicated as an intermediate in MMP-1 gene expression. However, the roles of individual NF-kappaB family members during IL-1beta-induced MMP-1 gene expression have not been defined.


RESULTS
To address the relationship between the NF-kappaB pathway and MMP-1 gene activation in chondrocytes, primary cultured human articular chondrocyte cultures (HAC) and SW-1353 cells were stimulated with IL-1beta over a 24-h time course and MMP-1, NF-kappaB1, NF-kappaB2 and RelA gene expression was assayed. IL-1beta-induced MMP-1 expression was comparable in HAC and SW-1353 cells both temporally and quantitatively. MMP-1 gene expression was mirrored by increases in NF-kappaB gene expression, and inhibition of NF-kappaB nuclear translocation with dominant-negative IkappaBalpha reduced IL-1beta-dependent MMP-1 gene expression. IL-1beta activated the NF-kappaB pathway in chondrocytes, both through phosphorylation and transient degradation of IkappaBalpha, as well as through sustained phosphorylation of RelA. Small inhibitory RNAs (siRNA) specific for RelA resulted in significant reduction of MMP-1 mRNA, whereas siRNA for NF-kappaB1 and NF-kappaB2 augmented IL-1beta-induced MMP-1 expression.


CONCLUSIONS
Our data demonstrate that IL-1beta activation of the NF-kappaB pathway is required for IL-1beta induction of MMP-1 in chondrocytes and that RelA can work independently of NF-kappaB1 or NF-kappaB2 to activate this gene expression program.
Raymond
L
L
Research Service, VA Medical Center, White River Junction, VT 05009, and Department of Medicine, Dartmouth Medical School, Lebanon, NH, USA.
Eck
S
S
Hays
E
E
Tomek
I
I
Kantor
S
S
Vincenti
M
M
D053674:Aggrecans / Q000502:physiology; D001168:Arthritis / Q000235:genetics*; D018929:Cell Culture Techniques / Q000379:methods; D019902:Chondrocytes / Q000502:physiology*; D024043:Collagen Type II / Q000502:physiology; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D053583:Interleukin-1beta / Q000235:genetics / Q000494:pharmacology*; D020781:Matrix Metalloproteinase 1 / Q000235:genetics*; D051996:Transcription Factor RelA / Q000235:genetics*; D015533:Transcriptional Activation
D053674:Aggrecans; D024043:Collagen Type II; D053583:Interleukin-1beta; D051996:Transcription Factor RelA; D020781:Matrix Metalloproteinase 1