Human hepatoma cells produce an 85 kDa gelatinase regulated by phorbol 12-myristate 13-acetate.
Several human cell lines were studied for the production of gelatinases. Diploid fibroblasts, the melanoma cell line Bowes, the MG-63 osteosarcoma cell line and the human hepatoma cell line Malavu all constitutively produced a 67 kDa gelatinase. Gelatinolytic enzymes were quantified by a sensitive zymographic substrate conversion assay. Upon induction with phorbol 12-myristate 13-acetate (PMA), the human hepatoma cell line secreted considerable amounts of an 85 kDa gelatinase activity. The induction process was time- and dose-dependent. It represented a true increase in production per individual cell and was associated by a marked change of the cell morphology. The effect of various proteinase inhibitors and the maximal activity of the enzyme near neutral pH demonstrate that it is a neutral metalloproteinase. Characterization studies showed the 85 kDa gelatinase to be transformed to lower molecular weight, active forms by treatment with p-aminophenylmercuric acetate (APMA) or trypsin.
Masure
S
S
Rega Institute for Medical Research, University of Leuven, Belgium.
Billiau
A
A
Van Damme
J
J
Opdenakker
G
G
D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D004789:Enzyme Activation; D018093:Gelatinases; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008113:Liver Neoplasms / Q000201:enzymology* / Q000473:pathology; D008666:Metalloendopeptidases / Q000378:metabolism; D008970:Molecular Weight; D010434:Pepsin A / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000201:enzymology
D011480:Protease Inhibitors; D010434:Pepsin A; D018093:Gelatinases; D008666:Metalloendopeptidases; D013755:Tetradecanoylphorbol Acetate