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J. Biol. Chem., Vol. 275, Issue 50, 39146-39151, December 15, 2000
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From the Kolling Institute of Medical Research, University of
Sydney, Royal North Shore Hospital, St. Leonards, New South Wales 2065, Australia
This study explores the relationship between
anti-proliferative signaling by transforming growth factor-
Growth Inhibition by Insulin-like Growth Factor-binding
Protein-3 in T47D Breast Cancer Cells Requires Transforming Growth
Factor-
(TGF-) and the Type II TGF- Receptor*
(TGF-) and insulin-like growth factor-binding protein-3 (IGFBP-3) in
human breast cancer cells. In MCF-7 cells, the expression of
recombinant IGFBP-3 inhibited proliferation and sensitized the cells to
further inhibition by TGF-1. To investigate the mechanism, we used
T47D cells that lack type II TGF- receptor (TGF-RII) and are
insensitive to TGF-1. After introducing the TGF-RII by
transfection, the basal proliferation rate was significantly decreased.
Exogenous TGF-1 caused no further growth inhibition, but
immunoneutralization of endogenous TGF-1 restored the proliferation
rate almost to the control level. The addition of IGFBP-3 did not
inhibit the proliferation of control cells but caused
dose-dependent inhibition in TGF-RII-expressing cells
when exogenous TGF-1 was also present. Similarly,
receptor-expressing cells showed dose-dependent sensitivity to exogenous TGF-1 only in the presence of exogenous IGFBP-3. This
indicates that in these cells, anti-proliferative signaling by
exogenous IGFBP-3 requires both the TGF-RII and exogenous TGF-1.
To investigate this synergism, the phosphorylation of TGF- signaling
intermediates, Smad2 and Smad3, was measured. Phosphorylation of each
Smad was stimulated by TGF-1 and, independently, by IGFBP-3 with the
two agents together showing a cumulative effect. These data suggest
that IGFBP-3 inhibitory signaling requires an active TGF- signaling
pathway and implicate Smad2 and Smad3 in IGFBP-3 signal transduction.
*
This study was supported by the University of Sydney Medical
Foundation.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.
To whom correspondence should be addressed. Tel.: 61-2-9926-8486;
Fax: 61-2-9926-8484; E-mail: robaxter@med.usyd.edu.au.
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