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J Biol Chem, Vol. 275, Issue 9, 6628-6635, March 3, 2000
From the Departments of Oncogenic ras induces the expression
of cyclooxygenase-2 (COX-2) in a variety of cells. Here we investigated
the role of transforming growth factor-
Transforming Growth Factor-
1 Enhances
Ha-ras-induced Expression of Cyclooxygenase-2 in Intestinal
Epithelial Cells via Stabilization of mRNA*
§,
,,§**
Surgery, ¶ Medicine, and
** Cell Biology, The Vanderbilt-Ingram Cancer Center, Vanderbilt
University Medical Center, Nashville, Tennessee 37232 and Eccles
Program in Human Molecular Biology and Genetics, Huntsman Cancer
Institute, University of Utah, Salt Lake City, Utah 84112
(TGF-) in the
Ras-mediated induction of COX-2 in intestinal epithelial cells (RIE-1).
RIE-1 cells were transfected with an inducible
Ha-RasVal12 cDNA and are referred as
RIE-iRas cells. the addition of 5 mM isopropyl-1-thio--D-galactopyranoside (IPTG) induced the
expression of Ha-RasVal12, closely followed by an increase
in the expression of COX-2. Neutralizing anti-TGF- antibody
partially blocked the Ras-induced increase in COX-2. Combined treatment
with IPTG and TGF-1 resulted in a 20-50-fold increase in the levels
of COX-2 mRNA. The t1/2 of COX-2 mRNA was
increased from 13 to 24 min by Ha-Ras induction alone. The addition of
TGF-1 further stabilized the COX-2 mRNA (t1/2 > 50 min). Stable transfection of a
luciferase reporter construct containing the COX-2 3'-untranslated region (3'-UTR) revealed that TGF-1 treatment and Ras induction each
stabilized the COX-2 3'-UTR. Combined treatment with IPTG and TGF-1
synergistically increased the luciferase activity. Furthermore, a
conserved AU-rich region located in the proximal COX-2 3'-UTR is
required for maximal stabilization of COX-2 3'-UTR by Ras or TGF-1
and is necessary for the synergistic stabilization of COX-2 3'-UTR by
oncogenic Ras and TGF-1.
*
This work was supported by National Institutes of Health
Grants DK-52334 and CA-69457 (to R. D. B.), DK-47297 and ES-00267 (to
R. N. D.), P01 CA77839 and CA 68485 (Vanderbilt Cancer Center), (to
R. D. B. and R. N. D.), and P01 CA73992 and CA42014 (to
S. M. P.).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 © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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