Induction of metalloproteinase activity in human T-lymphocytes.
Matrix metalloproteinases are thought to play major roles in a wide array of normal and pathological processes. These proteinases are involved in the degradation of the extracellular matrix and are believed to facilitate the movement of cells from one site to another. In the current study, we examined the expression of the 92 kDa gelatinase activity (MMP-9) by the human T-lymphoma cell line, HSB. Proteinase activity was greatly elevated when cells were treated with TPA. This induction was initially observed at 6 h post-TPA treatment and continued to increase up to 48 h. Proteinase induction was inhibited by actinomycin D and cycloheximide, indicating that nascent RNA and protein synthesis were required. Staurosporine, an inhibitor of protein kinase C activity, suppressed the TPA-induction of gelatinase activity. Our results suggest that TPA induces the 92 kDa gelatinase activity by activating protein kinase C. TGF-beta also induced proteinase activity, although to a lesser extent than TPA. Several criteria indicate that this enzyme is a member of the family of matrix metalloproteinases: (1) this activity was inhibited by EDTA, 1,10-phenanthroline and TIMP; (2) this activity bound to a gelatin-agarose affinity resin; (3) it has a mass of approx. 92 kDa on SDS-polyacrylamide gels; (4) it cleaves gelatin and (5) the inducible proteinase cross reacts with antiserum to MMP-9.
Department of Radiation Oncology, University of Pennsylvania School of Medicine, Philadelphia 19104.
H
H
Zhou
E J
EJ
Bernhard
F E
FE
Fox
P C
PC
Billings
D017364:Collagenases / Q000096:biosynthesis*; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D020780:Matrix Metalloproteinase 9; D061965:Matrix Metalloproteinase Inhibitors; D008666:Metalloendopeptidases / Q000096:biosynthesis*; D013601:T-Lymphocytes / Q000201:enzymology*; D013755:Tetradecanoylphorbol Acetate; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D061965:Matrix Metalloproteinase Inhibitors; D016212:Transforming Growth Factor beta; D003609:Dactinomycin; D003513:Cycloheximide; D017364:Collagenases; D008666:Metalloendopeptidases; D020780:Matrix Metalloproteinase 9; D013755:Tetradecanoylphorbol Acetate