title: Role of glucocorticoid receptor and CCAAT/enhancer-binding protein alpha in the feed-forward induction of 11beta-hydroxysteroid dehydrogenase type 1 expression by cortisol in human amnion fibroblasts.
abstract: The amount of cortisol available to its receptors is increased by the pre-receptor enzyme 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) which converts cortisone to cortisol. We examined the molecular mechanisms of the feedback effect of cortisol on 11beta-HSD1 mRNA expression in human amnion fibroblasts. Our data showed that cortisol-induced 11beta-HSD1 mRNA expression dose dependently in amnion fibroblasts, which could be completely blocked both by the mRNA transcription inhibitor 5,6-dichlorobenzimidazole riboside and by the glucocorticoid receptor (GR) antagonist RU486, and partially blocked by global inhibition of CCAAT/enhancer-binding proteins (C/EBPs) with transfection of C/EBP-specific dominant-negative expression CMV500 plasmid (AC/EBP) into the cells. Likewise, the induction of the promoter activity by cortisol could also be completely blocked by RU486 and partially by AC/EBP transfection. Progressive 5' deletion of the 11beta-HSD1promoter located the region responsible for cortisol's induction within -204 bp upstream to the transcription start site. Specific nucleotide mutations of the putative glucocorticoid responsive element or CCAAT in this promoter region attenuated the induction by cortisol. Moreover, chromatin immunoprecipitation assay and electrophoretic mobility shift assay showed that GR and C/EBPalpha but not C/EBPbeta could bind this promoter region upon cortisol stimulation of amnion fibroblasts. In conclusion, we demonstrated that GR and C/EBPalpha were involved in cortisol-induced 11beta-HSD1 mRNA expression via binding to 11beta-HSD1 promoter in amnion fibroblasts, which may cast a feed-forward production of cortisol in the fetal membranes at the end of gestation.
authors.affiliation: School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, People's Republic of China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yang
authors.forename: Chunming
authors.initials: C
authors.lastname: Guo
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.forename: Wenjiao
authors.initials: W
authors.lastname: Li
authors.forename: Leslie
authors.initials: L
authors.lastname: Myatt
authors.forename: Kang
authors.initials: K
authors.lastname: Sun
mesh_terms: D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1 / Q000096:biosynthesis* / Q000235:genetics; D000650:Amnion / Q000201:enzymology*; D001483:Base Sequence; D001562:Benzimidazoles / Q000494:pharmacology; D015153:Blotting, Western; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000502:physiology*; D002478:Cells, Cultured; D047369:Chromatin Immunoprecipitation; D004305:Dose-Response Relationship, Drug; D024202:Electrophoretic Mobility Shift Assay; D004790:Enzyme Induction / Q000187:drug effects; D025461:Feedback, Physiological; D005260:Female; D005347:Fibroblasts / Q000201:enzymology*; D005799:Genes, Dominant; D017930:Genes, Reporter; D005938:Glucocorticoids; D006727:Hormone Antagonists / Q000494:pharmacology; D006801:Humans; D006854:Hydrocortisone / Q000008:administration & dosage / Q000494:pharmacology*; D008156:Luciferases / Q000235:genetics; D015735:Mifepristone / Q000494:pharmacology; D009154:Mutation; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000502:physiology; D011965:Receptors, Glucocorticoid / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D020218:Response Elements / Q000235:genetics; D014162:Transfection
chemical_list: C110962:5,6-dichlorobenzimidazole; D001562:Benzimidazoles; D022763:CCAAT-Enhancer-Binding Protein-alpha; D005938:Glucocorticoids; D006727:Hormone Antagonists; D011965:Receptors, Glucocorticoid; D015735:Mifepristone; D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1; D008156:Luciferases; D006854:Hydrocortisone