Stimulation of MMP-11 (stromelysin-3) expression in mouse fibroblasts by cytokines, collagen and co-culture with human breast cancer cell lines.

Saxon Selvey,Michael G Irving,Larisa M Haupt,Erik W Thompson,Lyn R Griffiths,Klaus I Matthaei

doi:10.1186/1471-2407-4-40

Abstract

BackgroundMatrix metalloproteinases (MMPs) are central to degradation of the extracellular matrix and basement membrane during both normal and carcinogenic tissue remodeling. MT1-MMP (MMP-14) and stromelysin-3 (MMP-11) are two members of the MMP family of proteolytic enzymes that have been specifically implicated in breast cancer progression. Expressed in stromal fibroblasts adjacent to epithelial tumour cells, the mechanism of MT1-MMP and MMP-11 induction remains unknown.MethodsTo investigate possible mechanisms of induction, we examined the effects of a number of plausible regulatory agents and treatments that may physiologically influence MMP expression during tumour progression. Thus NIH3T3 and primary mouse embryonic fibroblasts (MEFs) were: a) treated with the cytokines IL-1β, IL-2, IL-6, IL-8 and TGF-β for 3, 6, 12, 24, and 48 hours; b) grown on collagens I, IV and V; c) treated with fibronectin, con-A and matrigel; and d) co-cultured with a range of HBC (human breast cancer) cell lines of varied invasive and metastatic potential.ResultsCompetitive quantitative RT-PCR indicated that MMP-11 expression was stimulated to a level greater than 100%, by 48 hour treatments of IL-1β, IL-2, TGF-β, fibronectin and collagen V. No other substantial changes in expression of MMP-11 or MT1-MMP in either tested fibroblast culture, under any treatment conditions, were observed.ConclusionWe have demonstrated significant MMP-11 stimulation in mouse fibroblasts using cytokines, matrix constituents and HBC cell lines, and also some inhibition of MT1-MMP. Our data suggest that the regulation of these genes in the complex stromal-epithelial interactions that occur in human breast carcinoma, is influenced by several mechanisms.

Highlights

Matrix metalloproteinases (MMPs) are central to degradation of the extracellular matrix and basement membrane during both normal and carcinogenic tissue remodeling
MMP-11 expression at 48 hours was affected by treatment with IL-1β, transforming growth factor (TGF)-β, fibronectin and collagen V, in the tested fibroblasts cultures
Similar to the MT1MMP results presented in the present study, MT1-MMP gene expression was reportedly not affected by insulinlike growth factor (IGF)-2, epidermal growth factor (EGF), platelet derived growth factor (PDGR)-BB, IL-6, TNF-β or IL-1β [28]

Summary

Introduction

Matrix metalloproteinases (MMPs) are central to degradation of the extracellular matrix and basement membrane during both normal and carcinogenic tissue remodeling. Expressed in stromal fibroblasts adjacent to epithelial tumour cells, the mechanism of MT1-MMP and MMP-11 induction remains unknown Integral to both normal and pathological tissue remodeling, the matrix metalloproteinase (MMP) family of proteolytic enzymes collectively degrades laminin, collagen, gelatin, and other protein components of the extracellular matrix [1,2]. MMP induction was measured in NIH3T3 and primary mouse embryonic fibroblasts (MEFs) subjected to each of the following potentially regulatory agents: collagen I, IV and V, concanavalin-A, fibronectin, and several exogenous cytokines. As induction by such agents could be transitory, treatment times were varied from 3, 6, 12, 24 and 48 hours, and were immediately followed by RNA extraction

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