Tumor-promoting phorbol esters and cell proliferation stimulate secretion of basement membrane (type IV) collagen-degrading metalloproteinase by human fibroblasts.
The secretion of a type IV collagen-specific proteinase is stimulated in cultured human skin fibroblasts by the phorbol ester tumor promoter 12-O-tetradecanoyl phorbol 13-acetate (TPA) and during cell proliferation. Exposure of the cells at the late log phase of growth to 10(-9) to 10(-6) M TPA resulted in the secretion of type IV collagenase activity to the medium, this effect being reversible. Incubation of intact type IV procollagen with TPA-induced fibroblast medium protein produced six peptides, four of which corresponded in size to the fragments produced by a type IV collagen-specific collagenase (Fessler, L., Duncan, K., Fessler, J., Salo, T., and Tryggvason (1984) J. Biol. Chem. 259, 9783-9789). The TPA-induced type IV collagen-degrading enzyme could be activated by trypsin, was inhibited by EDTA, but was not affected by soybean trypsin inhibitor, N-ethylmaleimide, aprotinin, or cysteine. Therefore, in human skin fibroblasts, TPA can induce a type IV collagen-specific, metal-dependent collagenase as was previously described in some invasive tumor cells. Furthermore, another metalloprotease is apparently secreted under the same conditions of TPA exposure. The production of metal-dependent, type IV collagen-degrading activity was also studied at different stages of cellular proliferation. In early log phase, a significant amount of enzyme activity was observed in the control cell medium; this activity disappeared during both late log and stationary growth phases. This activity could be markedly increased by the addition of 10(-8) M TPA to the culture medium. The production of matrix-degrading proteinases is therefore likely to be associated with rapid cell proliferation in both transformed and untransformed cells.
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T
Salo
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Turpeenniemi-Hujanen
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Tryggvason
D007611:Aprotinin / Q000494:pharmacology; D001485:Basement Membrane / Q000201:enzymology; D002455:Cell Division / Q000187:drug effects; D003470:Culture Media; D003545:Cysteine / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism*; D005033:Ethylmaleimide / Q000494:pharmacology; D005347:Fibroblasts / Q000201:enzymology / Q000648:ultrastructure; D006801:Humans; D008666:Metalloendopeptidases; D003012:Microbial Collagenase / Q000378:metabolism*; D010704:Phorbols / Q000494:pharmacology*; D011347:Procollagen / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000201:enzymology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism
D003470:Culture Media; D010704:Phorbols; D011347:Procollagen; D007611:Aprotinin; D004492:Edetic Acid; D010450:Endopeptidases; D014357:Trypsin; D008666:Metalloendopeptidases; D003012:Microbial Collagenase; D003545:Cysteine; D013755:Tetradecanoylphorbol Acetate; D005033:Ethylmaleimide