title: Activation of fibroblast-derived matrix metalloproteinase-2 by colon-cancer cells in non-contact Co-cultures.
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.
authors.affiliation: Department of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ko
authors.forename: S
authors.initials: S
authors.lastname: Yazumi
authors.forename: K
authors.initials: K
authors.lastname: Yoshikawa
authors.forename: Y
authors.initials: Y
authors.lastname: Konda
authors.forename: M
authors.initials: M
authors.lastname: Nakajima
authors.forename: T
authors.initials: T
authors.lastname: Chiba
authors.forename: R
authors.initials: R
authors.lastname: Takahashi
mesh_terms: D001485:Basement Membrane / Q000378:metabolism; D015152:Blotting, Northern; D018938:Caco-2 Cells; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D018920:Coculture Techniques; D003110:Colonic Neoplasms / Q000378:metabolism*; D004492:Edetic Acid / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005347:Fibroblasts / Q000201:enzymology*; D005780:Gelatin / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D011719:Pyrazines; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013449:Sulfonamides; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: C097658:CGS 27023A; D002614:Chelating Agents; D006877:Hydroxamic Acids; D011480:Protease Inhibitors; D011719:Pyrazines; D012333:RNA, Messenger; D013449:Sulfonamides; D019716:Tissue Inhibitor of Metalloproteinase-2; D005780:Gelatin; D004492:Edetic Acid; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2