Regulation of the promoter of the human corticotropin-releasing hormone gene in transfected human endometrial cells.
Corticotropin-releasing hormone (CRH) is expressed in several peripheral tissues, including normal epithelial cells of the human and rodent uterus. However, the biological role of endometrial CRH is known in neither species. As a first step to clarify this role, we studied the regulation of CRH promoter in endometrial cells. We performed homologous transfection experiments in Ishikawa cells, a human endometrial cell line, using a 0.9-kb fragment of the 5'-flanking region of the human CRH gene coupled to luciferase. Transfected cells were exposed for 18 h to 8-bromo cyclic adenosine monophosphate, forskolin, epidermal growth factor, steroids (estradiol, progesterone, and the synthetic glucocorticoid dexamethasone and their antagonists), and prostaglandin E2; then the activity of the luciferase reporter was determined in the cell lysates. We found that the activity of the 5'-flanking region of the CRH gene was stimulated by cyclic adenosine monophosphate and epidermal growth factor and inhibited in a receptor-mediated, dose-dependent fashion by estradiol and dexamethasone. The antiglucocorticoid RU 486 acted as a glucocorticoid agonist, suppressing the CRH gene activation, while progesterone was devoid of any activity. Prostaglandin E2 stimulated the CRH activation, and the prostanoid inhibitor indomethacin suppressed it, most probably by inhibiting endogenous prostaglandins. These findings suggest that endometrial CRH gene expression may be under the negative control of estrogens and glucocorticoids and under the positive control of prostaglandin E2.
Makrigiannakis
A
A
Department of Pharmacology, University of Crete Medical School, Iráklion, Greece.
Zoumakis
E
E
Margioris
A N
AN
Stournaras
C
C
Chrousos
G P
GP
Gravanis
A
A
D000230:Adenocarcinoma; D003346:Corticotropin-Releasing Hormone / Q000235:genetics*; D000242:Cyclic AMP / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D015232:Dinoprostone / Q000494:pharmacology; D016889:Endometrial Neoplasms; D004717:Endometrium / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics; D015735:Mifepristone / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins; D015533:Transcriptional Activation; D014162:Transfection; D014407:Tumor Cells, Cultured
D011993:Recombinant Fusion Proteins; D015735:Mifepristone; D004958:Estradiol; D004815:Epidermal Growth Factor; D003907:Dexamethasone; D003346:Corticotropin-Releasing Hormone; D000242:Cyclic AMP; D008156:Luciferases; D015232:Dinoprostone; D007213:Indomethacin