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Regulation of 92-kD gelatinase release in HL-60 leukemia cells: tumor
necrosis factor-alpha as an autocrine stimulus for basal- and phorbol
ester-induced secretion
C Ries, H Kolb and PE Petrides
Institute for Clinical Hematology, GSF Forschungszentrum fur Umwelt und
Gesundheit, Munich, Germany.
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 promyelocytic cell line HL-60 as determined by
zymographic analysis. Tumor necrosis factor-alpha (TNF-alpha) enhanced the
enzyme release threefold to fourfold and the protein kinase C (PKC)
activator and differentiation inducer 12-O-tetradecanoylphorbol-13- acetate
(TPA) eightfold to ninefold. Gelatinase induction by TNF-alpha and TPA was
inhibited by actinomycin D or cycloheximide, indicating that de novo
protein synthesis was required. Neutralizing monoclonal antibodies to
TNF-alpha (anti-TNF-alpha) 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-alpha expression in
HL-60 cells, such as pentoxifylline and dexamethasone, completely abrogated
both the constitutive and TPA-evoked MMP-9 release. Diethyldithiocarbamate,
which is known to stimulate TNF-alpha 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-alpha. High concentrations (2 mumol/L) of staurosporine completely
abolished the extracellular enzyme activity both in untreated and
TPA-stimulated cells. These results suggest, that TNF- alpha is required
for basal and PKC-mediated MMP-9 release in HL-60 leukemia cells. Thus,
MMP-9 secretion may be regulated by TNF-alpha 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.
Volume 83,
Issue 12,
pp. 3638-3646,
06/15/1994
Copyright © 1994 by The American Society of Hematology
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