Inhibition of proteolytic enzymes in the in vitro amnion model for basement membrane invasion.
The ability of B16-F10 mouse melanoma cells to cross an amnion basement membrane was determined in the presence of strong inhibitors of both serine and cysteine proteases. The concentrations of inhibitors were at orders of magnitude higher than their Ki values to serine and cysteine proteases implicated in metastasis, thus ensuring a complete inhibition for tumor secreted proteases such as cathepsin B-like proteases, plasminogen activators, and plasmin. Under these conditions of high serine and cysteine protease inhibitor concentrations, no significant decrease in B16-F10 melanoma cell invasion through the amnion was observed. Separate experiments showed that the inhibitors were neither toxic to the cells nor degraded. The results show that neither tumor cell secreted cathepsin B-like proteases nor plasminogen activator have a controlling role in basement membrane crossing in this metastatic model. A possible role for tumor cell membrane proteases in basement membrane invasion, in which the substrates of the protease bind to receptor sites near a membrane associated proteolytic activity, is not eliminated.
D000650:Amnion / Q000473:pathology; D000818:Animals; D001485:Basement Membrane / Q000473:pathology*; D001790:Blood Physiological Phenomena; D002401:Cathepsin B; D002403:Cathepsins / Q000502:physiology; D002462:Cell Membrane / Q000201:enzymology; D003546:Cysteine Endopeptidases; D010450:Endopeptidases; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007976:Leupeptins / Q000494:pharmacology; D008545:Melanoma / Q000473:pathology; D051379:Mice; D009361:Neoplasm Invasiveness; D010960:Plasminogen Activators / Q000502:physiology; D011247:Pregnancy; D011480:Protease Inhibitors / Q000494:pharmacology*; D012697:Serine Endopeptidases; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors
D007976:Leupeptins; D011480:Protease Inhibitors; D002403:Cathepsins; D010450:Endopeptidases; D010960:Plasminogen Activators; D012697:Serine Endopeptidases; D014568:Urokinase-Type Plasminogen Activator; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; C032854:leupeptin