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Activation of fibroblast‐derived matrix metalloproteinase‐2 by colon‐cancer cells in non‐contact co‐cultures
Author(s) -
Ko Kohaku,
Yazumi Shujiro,
Yoshikawa Kiyotsugu,
Konda Yoshitaka,
Nakajima Motowo,
Chiba Tsutomu,
Takahashi Rei
Publication year - 2000
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/1097-0215(20000715)87:2<165::aid-ijc2>3.0.co;2-f
Subject(s) - fibroblast , matrix metalloproteinase , cancer cell , cell culture , stromal cell , tissue inhibitor of metalloproteinase , cell , cancer research , biology , cancer , chemistry , zymography , microbiology and biotechnology , biochemistry , genetics
Abstract 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.