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J Biol Chem, Vol. 274, Issue 6, 3522-3530, February 5, 1999
, and
From the Division of Cardiology, University Hospital of Geneva,
Switzerland and the Angiotensin II (Ang II) and basic fibroblast
growth factor (bFGF) are important modulators of cell growth under
physiological and pathophysiological conditions. We and others have
previously shown that these growth factors increase insulin-like growth
factor-1 receptor (IGF-1R) number and mRNA in vascular smooth
muscle cells and that this effect is transcriptionally regulated. To
study the mechanisms and the signaling pathways involved, IGF-1R
promoter reporter constructs were transiently transfected in
CHO-AT1 cells that overexpress angiotensin
AT1 receptors. Our findings indicate that Ang II and bFGF
significantly increased IGF-1R promoter activity up to 7- and 3-fold,
respectively. The effect induced by Ang II was mediated via a tyrosine
kinase-dependent mechanism, since tyrphostin A25 largely
inhibited the Ang II-induced increase in promoter activity. In
addition, co-transfection of dominant negative Ras, Raf, and MEK1 or
pretreatment with the MEK inhibitor PD 98059 dose-dependently decreased both the Ang II- and
bFGF-induced increase in IGF-1R transcription and protein expression,
suggesting that the Ras-Raf-mitogen-activated protein kinase kinase
pathway is required for both growth factors. Reactive oxygen species
have been shown to act as second messengers in Ang II-induced
signaling, and activation of the transcription factor NF- Department of Medicine, Emory
University, Atlanta, Georgia 30322
B is
redox-sensitive. While co-transfection of dominant negative IB
mutant completely inhibited the Ang II-induced increase in
transcription, it had no effect on the bFGF signaling. In contrast,
co-transfection studies indicated that the transcription factors STAT1,
STAT3, and c-Jun and the Janus kinase 2 kinase are required in the
signaling pathway of bFGF, whereas only dominant c-Jun inhibited the
Ang II-induced effect. In summary, these data demonstrate that Ang II
and bFGF increase IGF-1R gene transcription via distinct as well as
shared pathways and have important implications for understanding growth-stimulatory effects of these growth factors on vascular cells.
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