Differential Effects of Transforming Growth Factor-β on the Expression of Collagenase-1 and Collagenase-3 in Human Fibroblasts*
- From the Departamento de Bioquı́mica y Biologı́a Molecular, Facultad de Medicina, Universidad de Oviedo, 33006 Oviedo, Spain
Abstract
Collagenase-3 (MMP-13) is a matrix metalloproteinase (MMP) originally identified in breast carcinomas which is also produced at significant levels during fetal ossification and in arthritic processes. In this work, we have found that transforming growth factor β1 (TGF-β1), a growth factor widely assumed to be inhibitory for MMPs, strongly induces collagenase-3 expression in human KMST fibroblasts. In contrast, this growth factor down-regulated the expression in these cells of collagenase-1 (MMP-1), an enzyme highly related to collagenase-3 in terms of structure and enzymatic properties. The positive effect of TGF-β1 on collagenase-3 expression was dose- and time-dependent, but independent of the effects of this growth factor on cell proliferation rate. Analysis of the signal transduction mechanisms underlying the up-regulating effect of TGF-β1 on collagenase-3 expression demonstrated that this growth factor acts through a signaling pathway involving protein kinase C and tyrosine kinase activities. Functional analysis of the collagenase-3 gene promoter region revealed that the inductive effect of TGF-β1 is partially mediated by an AP-1 site. Comparative analysis with the promoter region of the collagenase-1 gene which contains an AP-1 site at equivalent position, confirmed that TGF-β1 did not have any effect on CAT activity levels of this promoter. Finally, by using electrophoretic mobility shift assays and antibody supershift analysis, we propose that c-Fos, c-Jun, and JunD may play major roles in the collagenase-3 activation by TGF-β1 in human fibroblasts.
Footnotes
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↵* This work was supported in part by Grants SAF94-0892 and SAF97-0258 from the Comisión Interministerial de Ciencia y Tecnologı́a, EU-BIOMED II (BMH4-CT96-0017), and Glaxo-Wellcome, Spain.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.
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↵‡ Recipient of a fellowship from the Fundación para la Investigación Cientı́fica y Técnica (FICYT), Asturias, Spain.
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↵§ Recipient of a fellowship from the Ayuntamiento de Oviedo, Asturias, Spain.
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↵¶ To whom correspondence should be addressed: Departamento de Bioquı́mica y Biologı́a Molecular, Facultad de Medicina, Universidad de Oviedo, 33006 Oviedo, Spain. Tel.: 34-85-104201; Fax: 34-85-103564 or 34-85-232255; E-mail:clo{at}dwarf1.quimica.uniovi.es.
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↵1 The abbreviations used are: MMP, matrix metalloproteinase; IL-1β, interleukin 1β; TPA, 12-O-tetradecanoylphorbol-13-acetate; aFGF, acidic fibroblast growth factor; bFGF, basic fibroblast growth factor; TGF, transforming growth factor; CAT, chloramphenicol acetyltransferase; PKC, protein kinase C.
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↵2 J. A. Urı́a and C. López-Otı́n, unpublished results.
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- Received July 22, 1997.
- Revision received December 9, 1997.
- The American Society for Biochemistry and Molecular Biology, Inc.
