title: Steroid signalling in human ovarian surface epithelial cells: the response to interleukin-1alpha determined by microarray analysis.
abstract: The human ovarian surface epithelium (HOSE) is a common site of gynaecological disease including endometriosis and ovarian cancer, probably due to serial injury-repair events associated with successive ovulations. To comprehend the importance of steroid signalling in the regulation of the HOSE, we used a custom microarray to catalogue the expression of over 250 genes involved in the synthesis and reception of steroid hormones, sterols and retinoids. The array included a subset of non-steroidogenic genes commonly involved in pro-/anti-inflammatory signalling. HOSE cells donated by five patients undergoing surgery for non-malignant gynaecological conditions were cultured for 48 h in the presence and absence of 500 pg/ml interleukin-1alpha (IL-1alpha). Total RNA was reverse-transcribed into biotin-labelled cDNA, which was hybridised to the array and visualised by gold-particle resonance light scattering and charge-coupled device (CCD) camera detection. Results for selected genes were verified by quantitative reverse-transcription PCR. In five out of five cases, untreated HOSE cells expressed genes encoding enzymes required for de novo biosynthesis of cholesterol from acetate and subsequent formation of C21-pregnane and C19-androstane steroids. Consistent with the inability of HOSE cells to synthesise glucocorticoids, oestrogens or 5alpha-reduced androgens de novo, CYP21, CYP19 and 5alpha-reductase were not detected. The only steroidogenic gene significantly up-regulated by IL-1alpha was 11beta-hydroxysteroid dehydrogenase type 1 (11betaHSD1). Other cytokine-induced genes were IL-6, IL-8, nuclear factor kappaB (NFkappaB) inhibitor alpha, metallothionein-IIA and lysyl oxidase: inflammation-associated genes that respond to glucocorticoids. The only steroidogenic gene significantly suppressed by IL-1alpha was 3betaHSD1. Other genes suppressed by IL-1alpha were aldehyde dehydrogenase (ALDH) 1, ALDH 10, gonadotrophin hormone-releasing hormone receptor, peroxisome proliferation-activated receptor-binding protein (PPAR-bp) and nuclear receptor subfamily 2 group F member 2. These results define a steroidogenic phenotype of cultured HOSE cells and provide a limited expression profile for genes with associated signalling functions. IL-1alpha co-ordinately induces 11betaHSD1 and a panel of glucocorticoid-regulated, inflammation-associated genes in HOSE cells, providing further evidence that cortisol generated by 11betaHSD1 could participate in the local resolution of inflammation associated with ovulation.
authors.affiliation: University of Edinburgh Centre for Reproductive Biology, The Chancellor's Building, 49 Little France Crescent, Edinburgh EH16 4SB, UK. mrae1@staffmail.ed.ac.uk
authors.forename: M T
authors.initials: MT
authors.lastname: Rae
authors.forename: D
authors.initials: D
authors.lastname: Niven
authors.forename: A
authors.initials: A
authors.lastname: Ross
authors.forename: T
authors.initials: T
authors.lastname: Forster
authors.forename: R
authors.initials: R
authors.lastname: Lathe
authors.forename: H O D
authors.initials: HO
authors.lastname: Critchley
authors.forename: P
authors.initials: P
authors.lastname: Ghazal
authors.forename: S G
authors.initials: SG
authors.lastname: Hillier
mesh_terms: D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1 / Q000235:genetics; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology*; D007378:Interleukins / Q000235:genetics; D008668:Metallothionein / Q000235:genetics; D052002:NF-kappa B p50 Subunit; D020411:Oligonucleotide Array Sequence Analysis; D010053:Ovary / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D008249:Protein-Lysine 6-Oxidase / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology*; D013256:Steroids / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics
chemical_list: D007375:Interleukin-1; D007378:Interleukins; D052002:NF-kappa B p50 Subunit; C489548:NFKB1 protein, human; D013256:Steroids; D014157:Transcription Factors; D008668:Metallothionein; D043205:11-beta-Hydroxysteroid Dehydrogenase Type 1; D008249:Protein-Lysine 6-Oxidase