Human Co115 colon carcinoma cells potentiate the degradation of laminin mediated by tissue-type plasminogen activator.
The plasminogen activation (PA) system of human Co115 colon carcinoma cells was investigated. Analysis at the levels of protein and mRNA of cultured cells and of histozymography of tumor xenografts in nude mice showed that Co115 cells produce only tissue type PA (t-PA) and no urokinase (u-PA). Also, mRNA for the u-PA receptor and for PA inhibitor type 2 (PAI-2), but not for PAI-1, were detected. We developed a quantitative degradation assay using glutaraldehyde-immobilized 125I-laminin to investigate the capacity of Co115 cells to degrade laminin. Laminin degradation by Co115 cells was completely inhibited by 100 micrograms/ml of polyclonal anti-t-PA IgG, by the plasmin inhibitors aprotinin (100 micrograms/ml) or epsilon-aminocaproic acid (EACA; at 0.3 M), but not by antibodies against u-PA or u-PAR nor by nonimmune IgG. Cycloheximide-treated Co115 cells were unable to degrade laminin but increased laminin degradation induced by conditioned medium of Co115 cells or recombinant t-PA. No potentiation was observed when Co115 cells and laminin were kept separated by Transwell inserts. Our results suggest that Co115 human colon carcinoma cells degrade laminin by potentiating t-PA-mediated plasminogen activation at the cell surface which requires close contact between tumor cells and laminin substrate.
Swiss Institute for Experimental Cancer Research, Epalinges.
C
C
Tran-Thang
D
D
Vouillamoz
E K
EK
Kruithof
B
B
Sordat
D000818:Animals; D003110:Colonic Neoplasms / Q000473:pathology / Q000503:physiopathology*; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010960:Plasminogen Activators / Q000235:genetics / Q000502:physiology; D015849:Plasminogen Inactivators / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins; D010959:Tissue Plasminogen Activator / Q000378:metabolism*; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured
D007797:Laminin; C524898:PLAUR protein, human; D015849:Plasminogen Inactivators; C524899:Plaur protein, mouse; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins; D010960:Plasminogen Activators; D010959:Tissue Plasminogen Activator