Activation of fibroblast-derived matrix metalloproteinase-2 by colon-cancer cells in non-contact Co-cultures

Int J Cancer. 2000 Jul 15;87(2):165-71.


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.

MeSH terms

  • Basement Membrane / metabolism
  • Blotting, Northern
  • Caco-2 Cells
  • Cell Line
  • Cell Membrane / metabolism
  • Chelating Agents / pharmacology
  • Coculture Techniques
  • Colonic Neoplasms / metabolism*
  • Edetic Acid / pharmacology
  • Enzyme Activation / drug effects
  • Fibroblasts / enzymology*
  • Gelatin / metabolism
  • Humans
  • Hydroxamic Acids*
  • Matrix Metalloproteinase 2 / metabolism*
  • Matrix Metalloproteinases, Membrane-Associated
  • Metalloendopeptidases / metabolism
  • Protease Inhibitors / pharmacology
  • Pyrazines*
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sulfonamides
  • Tissue Inhibitor of Metalloproteinase-2 / pharmacology
  • Tumor Cells, Cultured


  • CGS 27023A
  • Chelating Agents
  • Hydroxamic Acids
  • Protease Inhibitors
  • Pyrazines
  • RNA, Messenger
  • Sulfonamides
  • Tissue Inhibitor of Metalloproteinase-2
  • Gelatin
  • Edetic Acid
  • Matrix Metalloproteinases, Membrane-Associated
  • Metalloendopeptidases
  • Matrix Metalloproteinase 2