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J Biol Chem, Vol. 273, Issue 14, 7800-7806, April 3, 1998
Gene Is Transcriptionally
Activated by Epidermal Growth Factor through cAMP Response Element
in Trophoblast Cells
From the Department of Obstetrics and Gynecology, Osaka University
Medical School, Suita, Osaka 565, Japan
The purpose of this study was to analyze the
mechanism of transcriptional activation of human chorionic
gonadotropin-
(hCG) gene by epidermal growth factor (EGF) in
trophoblast cells. We stably transfected hCG
promoter-chloramphenicol acetyltransferase constructs into Rcho-1
trophoblast cells and monitored the promoter activities. 
290-base
pair hCG promoter containing a tandem repeat of cAMP response
element (CRE) was activated by EGF in a dose- and
time-dependent manner. Deletion analysis of hCG promoter suggested an involvement of CRE in EGF-induced hCG transcriptional activation. Moreover, the hCG promoter, of which both CREs were mutated, did not respond to EGF. These results indicate that EGF activates the hCG gene transcription through CRE. Although EGF did
not alter the amount of CRE-binding protein (CREB), EGF induced CREB
phosphorylation. We next examined the mechanism of CREB phosphorylation by EGF. Protein kinase C inhibitors (H7, staurosporin, and
chelerythrine) inhibited EGF-induced CREB phosphorylation, whereas
either mitogen-activated protein kinase kinase-1 inhibitor (PD98059) or
protein kinase A inhibitor (H8) showed no effect. Furthermore, H7 and
staurosporin but not H8 inhibited hCG promoter activation by EGF. In
conclusion, EGF promotes hCG gene transcription via the CRE region
probably by phosphorylating CREB mainly through the protein kinase C
pathway in trophoblast cells.
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