Activation of fibroblast-derived matrix metalloproteinase-2 by colon cancer cells in noncontact co-cultures

Stromal fibroblasts interact with invading cancer cells by secreting and activating matrix metalloproteinases (MMPs). To elucidate the mechanisms involved in the expression and activation patterns of MMPs, human colon-cancer cell lines Caco-2 and LoVo and colon-fibroblast cell line CCD18-Co were co-cultivated in non-contact and contact conditions which mimic in vivo interaction between cancer cells and fibroblasts before and after cancer invasion respectively. Gelatin zymography disclosed that MMP-2 was secreted from the fibroblasts but not from the cancer cells. The quantity of fibroblast-derived MMP-2 in conditioned medium was not significantly changed in either the contact or the non-contact co-cultures when compared with that of individual cultures of CCD18-Co fibroblasts. Cancer cells in non-contact co-cultures, however, enhanced the activation of fibroblast-derived MMP-2. Transcripts of membrane-type matrix metalloproteinase-1 (MT1-MMP), which is thought to be present on the cell surface and to work as a candidate activator of MMP-2, were detected in both cancer cell lines. Plasma membrane extracts of cancer cells also activated MMP-2 in conditioned media in cell-free conditions. This activation of MMP-2 may be caused by MT1-MMP of the cancer cells, since it was inhibited by a series of MMP inhibitors, including ethylenediaminetetraacetic acid (EDTA), the tissue inhibitor of metalloproteinase-2 (TIMP-2), and the MMP inhibitor CGS 27023A, but not by TIMP-1. Our data demonstrate that in non-contact co-cultures colon-cancer cells activate fibroblast-derived MMP-2 on their plasma membranes. These findings should help to elucidate the mechanism involved in the initial destruction of basement membrane by cancer cells. Int. J. Cancer 87:165–171, 2000. © 2000 Wiley-Liss, Inc.