Stimulation of MMP-II (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. MTI-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 MTI-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-Ibeta, 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-II expression was stimulated to a level greater than 100%, by 48 hour treatments of IL-Ibeta, IL-2, TGF-beta, fibronectin and collagen V. No other substantial changes in expression of MMP-II or MTI-MMP in either tested fibroblast culture, under any treatment conditions, were observed. Conclusion: We have demonstrated significant MMP-II stimulation in mouse fibroblasts using cytokines, matrix constituents and HBC cell lines, and also some inhibition of MTI-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.