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J. Biol. Chem., Vol. 277, Issue 17, 14829-14837, April 26, 2002
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From the 12-O-Tetradecanoylphorbol-13-acetate
(TPA) suppresses the proliferation of the human breast epithelial cell
line MCF10A-Neo by initiating proteolytic processes that activate
latent transforming growth factor (TGF)-
Phorbol Ester Activation of a Proteolytic Cascade Capable of
Activating Latent Transforming Growth Factor-
A PROCESS INITIATED BY THE EXOCYTOSIS OF CATHEPSIN B*
,,¶
Institute of Environmental Health Sciences,
Wayne State University and the § Department of Pharmacology,
Wayne State University School of Medicine,
Detroit, Michigan 48201
in the serum used to
supplement culture medium. Within 1 h of treatment, cultures
accumulated an extracellular activity capable of cleaving a substrate
for urokinase-type plasminogen activator (uPA) and tissue plasminogen
activator (tPA). This activity was inhibited by plasminogen activator
inhibitor-1 or antibodies to uPA but not tPA. Pro-uPA activation was
preceded by dramatic changes in lysosome trafficking and the
extracellular appearance of cathepsin B and -hexosaminidase but not
cathepsins D or L. Co-treatment of cultures with the cathepsin B
inhibitors CA-074 or Z-FA-FMK suppressed the cytostatic effects of TPA
and activation of pro-uPA. In the absence of TPA, exogenously added
cathepsin B activated pro-uPA and suppressed MCF10A-Neo proliferation.
The cytostatic effects of both TPA and cathepsin B were suppressed in
cells cultured in medium depleted of plasminogen/plasmin or supplemented with neutralizing TGF- antibody. Pretreatment with cycloheximide did not suppress the exocytosis of cathepsin B or the
activation of pro-uPA. Hence, TPA activates signaling processes that
trigger the exocytosis of a subpopulation of lysosomes/endosomes containing cathepsin B. Subsequently, extracellular cathepsin B
initiates a proteolytic cascade involving uPA, plasminogen, and plasmin
that activates serum-derived latent TGF-.
*
This work was supported in part by National Institutes of
Health Grants CA34469 and CA56586 and was assisted by the services of
the Cell Culture Core and Cell Imaging and Cytometry Facility Core,
which is supported by National Institutes of Environmental Health
Sciences Grant P30 ES06639.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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