Insensitivity of laminin degradation directed by receptor bound urokinase to PAI-1 in cultured colon cancer.
This laboratory recently reported that laminin degradation by cultured colon cancer was plasminogen dependent and reflected the presence of urokinase bound to cell surface receptors. (Schlecte, W.; Murano, G.; Boyd D. Cancer Res., 49:6064-6069; 1989). The present study was undertaken to determine the sensitivity of urokinase receptor directed proteolysis to the type I plasminogen activator inhibitor (PAI-1). Colon cancer cell types, that were highly effective in degrading laminin in vitro, elaborated into their conditioned medium an inhibitor which was indistinguishable from PAI-1 on the basis of its performance in reverse zymography, western blotting, and immunoprecipitation assays. A fraction of this PAI-1 was active, as evidenced by complex formation with the active site of radioactive urokinase. Laminin degradation by the colon cancer cells, however, did not appear to be affected by the endogenous inhibitor, since an antibody to the inhibitor, which blocked urokinase-PAI-1 interactions, had little effect on laminin turnover. Further, addition of exogenous PAI-1, activated by guanidine hydrochloride pretreatment, to the colon cancer cells did not perturb laminin degradation. Because laminin degradation by colonic cells was a function of receptor bound urokinase, presumably immobilized plasminogen activator escaped the neutralizing effect of the inhibitor. These data suggest either a shielding effect of the receptor on the plasminogen activator or a physical separation of activator and inhibitor. Either way, for cultured colon cancer at least, laminin degradation directed by urokinase receptor bound plasminogen activator appeared unaffected by the presence of this inhibitor.
Schlechte
J W
JW
Bristol-Baylor Laboratory, Pharmacology Department, Baylor College of Medicine, Houston, TX 77030.
Boyd
D D
DD
D015153:Blotting, Western; D002478:Cells, Cultured; D003110:Colonic Neoplasms / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D066298:In Vitro Techniques; D007797:Laminin / Q000378:metabolism*; D015849:Plasminogen Inactivators / Q000302:isolation & purification / Q000494:pharmacology*; D011233:Precipitin Tests; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D007797:Laminin; D015849:Plasminogen Inactivators; D014568:Urokinase-Type Plasminogen Activator