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


METHODS
To 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-1beta, IL-2, IL-6, IL-8 and TGF-beta 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.


RESULTS
Competitive quantitative RT-PCR indicated that MMP-11 expression was stimulated to a level greater than 100%, by 48 hour treatments of IL-1beta, IL-2, TGF-beta, 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.


CONCLUSIONS
We 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.
Selvey
Saxon
S
Genomics Research Center, School of Heath Science, Griffith University, Gold Coast, Australia. l.griffiths@griffith.edu.au
Haupt
Larisa M
LM
Thompson
Erik W
EW
Matthaei
Klaus I
KI
Irving
Michael G
MG
Griffiths
Lyn R
LR
D016475:3T3 Cells; D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism; D003094:Collagen / Q000378:metabolism* / Q000494:pharmacology; D016207:Cytokines / Q000378:metabolism*; D004338:Drug Combinations; D005347:Fibroblasts / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D053507:Matrix Metalloproteinase 11; D053511:Matrix Metalloproteinase 14; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D011509:Proteoglycans / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D016207:Cytokines; D004338:Drug Combinations; D005353:Fibronectins; D007797:Laminin; C507096:Mmp14 protein, mouse; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D053507:Matrix Metalloproteinase 11; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D053511:Matrix Metalloproteinase 14