Steroid signalling in human ovarian surface epithelial cells: the response to interleukin-1alpha determined by microarray analysis

M T Rae, D Niven, A Ross, T Forster, R Lathe, H O D Critchley, P Ghazal, S G Hillier

Research output: Contribution to journalArticlepeer-review

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.
Original languageEnglish
Pages (from-to)19-28
Number of pages10
JournalJournal of Endocrinology
Volume183
Issue number1
DOIs
Publication statusPublished - 2004

Keywords

  • 11-beta-Hydroxysteroid Dehydrogenase Type 1
  • Cells, Cultured
  • Epithelial Cells
  • Female
  • Gene Expression
  • Gene Expression Profiling
  • Humans
  • Interleukin-1
  • Interleukins
  • Metallothionein
  • NF-kappa B p50 Subunit
  • Oligonucleotide Array Sequence Analysis
  • Ovary
  • Protein-Lysine 6-Oxidase
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Steroids
  • Transcription Factors

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