Induction of Collagenase-3 (MMP-13) Expression in Human Skin Fibroblasts by Three-dimensional Collagen Is Mediated by p38 Mitogen-activated Protein Kinase

doi:10.1074/jbc.274.4.2446

Induction of Collagenase-3 (MMP-13) Expression in Human Skin Fibroblasts by Three-dimensional Collagen Is Mediated by p38 Mitogen-activated Protein Kinase*

From the ^‡Department of Dermatology, Turku University Central Hospital and the ^§Department of Medical Biochemistry, and MediCity Research Laboratory, University of Turku, FIN-20520 Turku, Finland, the ^¶Department of Biological and Environmental Science, University of Jyväskylä, FIN-40351 Jyväskylä, Finland, and the ^‖Departamento Bioquı́mica y Biologı́a Molecular, Universidad de Oviedo, 33006 Oviedo, Spain

Abstract

Collagenase-3 (matrix metalloproteinase-13, MMP-13) is a recently identified human MMP with an exceptionally wide substrate specificity and restricted tissue-specific expression. Here we show that MMP-13 expression is induced in normal human skin fibroblasts cultured within three-dimensional collagen gel resulting in production and proteolytic activation of MMP-13. Induction of MMP-13 mRNAs by collagen gel was potently inhibited by blocking antibodies against α₁ and α₂ integrin subunits and augmented by activating antibody against β₁ integrin subunit, indicating that both α₁β₁ and α₂β₁ integrins mediate the MMP-13-inducing cellular signal generated by three-dimensional collagen. Collagen-related induction of MMP-13 expression was dependent on tyrosine kinase activity, as it was abolished by treatment of fibroblasts with tyrosine kinase inhibitors genistein and herbimycin A. Contact of fibroblasts to three-dimensional collagen resulted in simultaneous activation of mitogen-activated protein kinases (MAPKs) in three distinct subgroups: extracellular signal-regulated kinase (ERK)1 and ERK2, Jun N-terminal kinase/stress-activated protein kinase, and p38. Induction of MMP-13 expression was inhibited by treatment of fibroblasts with a specific p38 inhibitor, SB 203580, whereas blocking the ERK1,2 pathway (Raf/MEK1,2/ERK1,2) by PD 98059, a selective inhibitor of MEK1,2 activation potently augmented MMP-13 expression. Furthermore, specific activation of ERK1,2 pathway by 12-O-tetradecanoylphorbol-13-acetate markedly suppressed MMP-13 expression in dermal fibroblasts in collagen gel. These results show that collagen-dependent induction of MMP-13 in dermal fibroblasts requires p38 activity, and is inhibited by activation of ERK1,2. Therefore, the balance between the activity of ERK1,2 and p38 MAPK pathways appears to be crucial in regulation of MMP-13 expression in dermal fibroblasts, suggesting that p38 MAPK may serve as a target for selective inhibition of collagen degradation, e.g. in chronic dermal ulcers.

Footnotes

↵* This work was supported by grants from the Academy of Finland, the Sigrid Jusélius Foundation, the Cancer Research Foundation of Finland, Turku University Central Hospital, and the Turku University Foundation.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵** To whom correspondence should be addressed: MediCity Research Laboratory, University of Turku, Tykistökatu 6A, FIN-20520 Turku, Finland. Tel.: 358-2-3337025; Fax: 358-2-3337000; E-mail:veli-matti.kahari{at}utu.fi.
Abbreviations:

ECM

extracellular matrix

MMP

matrix metalloproteinase

TNF-α

tumor necrosis factor-α

IL-1

interleukin-1

TGF-β

transforming growth factor-β

MAPK

mitogen-activated protein kinase

ERK

extracellular signal-regulated kinase

JNK/SAPK

Jun N-terminal kinase/stress-activated protein kinase

MEK

MAPK/ERK kinase

TPA

12-O-tetradecanoylphorbol-13-acetate

PKC

protein kinase C

SCC

squamous cell carcinoma

FAK

focal adhesion kinase

RT

reverse transcription

PCR

polymerase chain reaction

GAPDH

glyceraldehyde-3-phosphate dehydrogenase

mAb

monoclonal antibody

bp

base pair(s)

kb

kilobase pair(s)

DMEM

Dulbecco’s modified Eagle’s medium

FCS

fetal calf serum.
- Received April 24, 1998.
- Revision received September 18, 1998.
The American Society for Biochemistry and Molecular Biology, Inc.