Secretion of proteinase inhibitors by tumorigenic and nontumorigenic guinea pig and Syrian hamster fibroblasts: evidence for autocrine regulation of local proteolysis.
A cytokine that inhibits fibrinolysis has been detected in the serum-free culture medium of guinea pig and hamster fibroblasts. This proteinase inhibitor was also present in Triton X-100 extracts of guinea pig cells. It was stable at pH 3.0 for 2 hr and was produced by cells rather than assimilated from serum in the culture medium as evidenced by: (a) an apparent molecular size (less than 45 kilodaltons) less than that of the principal serum-derived proteinase inhibitors; (b) its continued secretion after several passages of the cells in serum-free medium; and (c) the lack of inhibitory activity in the medium of mitomycin C-treated cells. The cytokine inhibited the proteinase activity of human urokinase, soluble TPA-stimulated guinea pig plasminogen activator, and the cell-associated plasminogen activator of tumorigenic guinea pig cells. Soluble plasminogen activator appeared to be inhibited to a greater degree than the cell-associated enzyme. The fluorogenic substrate (7-(N-carbobenzoxyglycylglycylargininamido)-4-methylcoumarin was used in a direct assay of proteinase activity and demonstrated that the cytokine inhibited both plasminogen activator and plasmin, the two proteinases of the fibrinolytic cascade. Tumorigenic guinea pig and hamster fibroblasts as well as nontransformed guinea pig fibroblasts were found to produce the inhibitory cytokine, and the amount of inhibitor secreted was independent of the tumorigenic potential of the cells. Production of the inhibitor by normal cells may be related to contact inhibition of growth, and this cytokine may contribute to the fine regulation of local proteolysis within tissues.
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000633:toxicity; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D005342:Fibrinolysis; D005347:Fibroblasts / Q000378:metabolism; D006168:Guinea Pigs; D008647:Mesocricetus; D016685:Mitomycin; D008937:Mitomycins / Q000633:toxicity; D008970:Molecular Weight; D009374:Neoplasms, Experimental / Q000378:metabolism*; D011480:Protease Inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
D000903:Antibiotics, Antineoplastic; D008937:Mitomycins; D011480:Protease Inhibitors; D016685:Mitomycin; D014568:Urokinase-Type Plasminogen Activator