Enhancement of Fibroblast Collagenase (Matrix Metalloproteinase-1)Gene Expression by Ceramide Is Mediated by Extracellular Signal-regulated and Stress-activated Protein Kinase Pathways

Abstract

Inflammatory cytokines tumor necrosis factor-␣ and interleukin-1 trigger the ceramide signaling pathway, initiated by neutral sphingomyelinase-elicited hydrolysis of cell membrane phospholipid sphingomyelin to ceramide, a new lipid second messenger. Here, we show that triggering the ceramide pathway by sphingomyelinase or C 2 -and C 6 -ceramide enhances collagenase-1 (matrix metalloproteinase-1; MMP-1) gene expression by fibroblasts. C 2 -ceramide activates three distinct mitogen-activated protein kinases (MAPKs) in dermal fibroblasts, i.e. extracellular signal-regulated kinase 1/2 (ERK1/2), stress-activated protein kinase/Jun N-terminal-kinase (SAPK/JNK), and p38. Stimulation of MMP-1 promoter activity by C 2 -ceramide is dependent on the presence of a functional AP-1 cis-element and is entirely inhibited by overexpression of MAPK inhibitor, dual specificity phosphatase CL100 (MAPK phosphatase-1). Activation of MMP-1 promoter by C 2 -ceramide is also effectively inhibited by kinase-deficient forms of ERK1/2 kinase (MEK1/2) activator Raf-1, ERK1 and ERK2, SAPK/ JNK activator SEK1, or SAPK␤. In addition, ceramidedependent induction of MMP-1 expression is potently prevented by PD 98059, a selective inhibitor of MEK1 activation, and by specific p38 inhibitor SB 203580. These results show that triggering the ceramide signaling pathway activates MMP-1 gene expression via three distinct MAPK pathways, i.e. ERK1/2, SAPK/JNK, and p38, and suggest that targeted modulation of the ceramide signaling pathway may offer a novel therapeutic approach for inhibiting collagenolytic activity, e.g. in inflammatory disorders.

Inhibition of collagenase-3 (MMP-13) expression in transformed human keratinocytes by interferon-γ is associated with activation of extracellular signal-regulated kinase-1,2 and STAT1

“…This observation is in agreement with recent studies with PD98059, demonstrating that ERK1,2 pathway mediates activation of MMP-1 expression in ®broblasts (Reunanen et al, 1998;Westermarck et al, 1998) and MMP-9 expression in transformed keratinocytes (Gum et al, 1997). Interestingly, a recent study demonstrated that IFN-g inhibits TGF-b signaling in ®brosarcoma cells by inducing expression of SMAD7, an inhibitory member of the SMAD transcription factor family (Ulloa et al, 1999).…”

“…In addition, overexpression of tissue inhibitor of metalloproteinases (TIMP)-1, -2, and -3 appears an e ective way of inhibiting the invasion of malignant cells . Our results together with recent observations (Reunanen et al, 1998;Ravanti et al, 1999;Westermarck et al, 1998;Gum et al, 1997) indicate that modulation of the activity of distinct MAPKs may serve as a potent way of inhibiting MMP expression and invasion. In conclusion, the results of the present study show that selective activation of the IFN-g signaling pathway may serve as a speci®c way of potently suppressing invasion and metastasis of malignant squamous epithelial cells in vivo.…”

“…Western blot analysis was performed as previously described (Reunanen et al, 1998) with antibodies speci®c for Tyr-701-phosphorylated STAT1, Ser-727-phosphorylated STAT1, ERK1,2, JNK/SAPK, and p38 in dilution 1 : 1000. Speci®c binding of antibodies was detected with peroxidase conjugated secondary antibodies and visualized by ECL.…”

c-Jun N-terminal kinase is required for metalloproteinase expression and joint destruction in inflammatory arthritis

“…For instance, p38 might play an important role in phorbol ester-induced type IV collagenase production by a squamous cell carcinoma cell line as well as in cytokine-stimulated chondrocytes (27,28). ERK and JNK have been implicated in the regulation of collagenase gene expression in cultured fibroblasts (29). In contrast to FLS, collagenase and stromelysin gene expression in endothelial cells are dependent on p38 (30).…”

Matrix metalloproteinase‐19 is expressed by proliferating epithelium but disappears with neoplastic dedifferentiation

“…23 Northern blot hybridizations were performed as described previously 24 with cDNAs labeled with [␣-32 P]-dCTP using random priming. A 1.5 kb cDNA was used for MMP-19 7 detection.…”