Tl. Mccarthy et al., 17-BETA-ESTRADIOL POTENTLY SUPPRESSES CAMP-INDUCED INSULIN-LIKE GROWTH-FACTOR-I GENE ACTIVATION IN PRIMARY RAT OSTEOBLAST CULTURES, The Journal of biological chemistry, 272(29), 1997, pp. 18132-18139
Insulin-like growth factor-I (IGF-I) is a key factor in bone remodelin
g. In osteoblasts, IGF-I synthesis is enhanced by parathyroid hormone
and prostaglandin E-2 (PGE(2)) through cAMP-activated protein kinase.
In rats, estrogen loss after ovariectomy leads to a rise in serum IGF-
I and an increase in bone remodeling, both of which are reversed by es
trogen treatment, To examine estrogen-dependent regulation of IGF-P ex
pression at the molecular level, primary fetal rat osteoblasts were co
-transfected with the estrogen receptor (hER, to ensure active ER expr
ession), and luciferase reporter plasmids controlled by promoter I of
the rat IGF-I gene (HGF-P PP), used exclusively in these cells, Bs rep
orted, 1 mu M PGE(2) increased IGF-I P1 activity by 5-fold. 17 beta-Es
tradiol alone had no effect, but dose-dependently suppressed the stimu
latory effect of PGE(2) by up to 90% (ED50 similar to-0.1 nM). This oc
curred within 3 In, persisted for at least 16 h, required ER, and appe
ared specific, since 17 alpha-estradiol was 100-300-fold less effectiv
e, By contrast, 17 beta-Estradiol stimulated estrogen response element
(ERE)dependent reporter expression by up to 10-fold, 17 beta-Estradio
l also suppressed an HGF-I PP. construct retaining only minimal promot
er sequence required for cAMP-dependent gene activation, but did not a
ffect the 60-fold increase in cAMP induced by PGE(2). There is no cons
ensus ERE in rat HGF-H P1, suggesting novel downstream interactions in
the cAMP pathway that normally enhances IGF-I expression ist skeletal
cells, To explore this, nuclear extract from osteoblasts expressing h
ER were examined by electrophoretic mobility shift assay using the aty
pical cAMP response element in IGF-I P1. Estrogen alone did not cause
DNA-protein binding, while PGE(2) induced a characteristic gel shift c
omplex. eo-treatment with both hormones caused a gel shift greatly dim
inished in intensity, consistent with their combined effects on IGF-H
promoter activity. Nonetheless, hER did not bind IGF-I cAMP response e
lement or any adjacent sequences,These results provide new molecular e
vidence that estrogen may temper the biological effects of hormones ac
ting through cAMP to regulate skeletal IGF-I expression and activity,.