Regulation of 92-kD Gelatinase Release in HL-60 Leukemia Cells: Tumor Necrosis Factor-α as an Autocrine Stimulus for Basal- and Phorbol Ester-Induced Secretion

• Published in: Blood Vol. 83; no. 12; pp. 3638 - 3646
• Main Authors: Ries, Christian, Kolb, Helmut, Petrides, Petro E.
• Format: Journal Article
• Language: English
• Published: Washington, DC Elsevier Inc 15.06.1994; The Americain Society of Hematology
• Subjects: Alkaloids - pharmacology; Biological and medical sciences; Dexamethasone - pharmacology; Ditiocarb - pharmacology; Gelatinases - metabolism; Hematologic and hematopoietic diseases; Humans; Leukemia, Promyelocytic, Acute - enzymology; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis; Medical sciences; Molecular Weight; Pentoxifylline - pharmacology; Protein Kinase C - physiology; Staurosporine; Tetradecanoylphorbol Acetate - pharmacology; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha - pharmacology; Tumor Necrosis Factor-alpha - physiology; Human; Promyelocytic leukemia; Cell culture; Enzyme; Acute; Cytokine; Metalloendopeptidases; Exploration; Malignant hemopathy; Metalloproteins; Gelatinase A; Regulation(control); Cell line; Enzymatic activity; Hydrolases; Proteinases; Tumor necrosis factor α
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V83.12.3638.3638

Matrix metalloproteinase 9 (MMP-9), also known as 92-kD type IV collagenase/gelatinase, is believed to play a critical role in tumor invasion and metastasis. Here, we report that MMP-9 was constitutively released from the human promy-elocytic cell line HL-60 as determined by zymographic analysis. Tumor necrosis factor-α (TNF-α) enhanced the enzyme release threefold to fourfold and the protein kinase C (PKC) activator and differentiation inducer 12-O-tetradecanoyl-phorbol-13-acetate (TPA) eightfold to ninefold. Gelatinase induction by TNF-α and TPA was inhibited by actinomycin D or cycloheximide, indicating that de novo protein synthesis was required. Neutralizing monoclonal antibodies to TNF-α (anti-TNF-α) decreased the basal MMP-9 release of these cells. In addition, these antibodies also significantly interfered with the TPA-induced enzyme release. Agents that inhibit TNF-α expression in HL-60 cells, such as pentoxifylline and dexamethasone, completely abrogated both the constitutive and TPA-evoked MMP-9 release. Diethyldithio-carbamate, which is known to stimulate TNF-α production in HL-60 cells, exerted a positive effect on MMP-9 release in untreated cells but was inhibitory in TPA-treated HL-60 cells. The PKC inhibitor staurosporine at low concentrations (100 ng/mL) caused a significant augmentation of MMP-9 release in untreated cultures that was blocked by the addition of anti-TNF-α. High concentrations (2 μmol/L) of staurosporine completely abolished the extracellular enzyme activity both in untreated and TPA-stimulated cells. These results suggest, that TNF-α is required for basal and PKC-mediated MMP-9 release in HL-60 leukemia cells. Thus, MMP-9 secretion may be regulated by TNF-α not only in a paracrine but also in an autocrine fashion. This may potentiate the matrix degradative capacity of immature leukemic cells in the processes of bone marrow egress and the evasion of these cells into peripheral tissue.