Ovulation involves rupture of the ovarian surface, and thus the ovarian surface epithelium (OSE) is subjected to serial injury and repair with each ovulatory cycle. Inflammation is integral to the process of ovulation, but importantly, inflammatory damage is also thought to be a potential mechanism of ovarian tumour development. We have examined OSE responses to an inflammatory stimulus associated with ovulation, Interleukin-1alpha (IL1alpha), using a custom designed steroid signalling oligonucletide microarray in combination with a sensitive detection platform suitable for use with the limited amounts of RNA recoverable from primary cell cultures.
Human OSE were collected with informed consent and local ethical committee approval from 5 women undergoing laparotomy for benign gynaecological conditions. Cells were cultured for 3 weeks, then, treated plus/minus 500 picogrammes per millilitre IL1alpha for 48 hours. RNA was extracted and reverse transcribed; cDNA was then used to probe the customised oligonucleotide gene array.
Genes involved in mediation of inflammation such as Metallothionin-IIA, Interleukin-6 and 8 were up-regulated by IL1a treatment (P<0.01). Lysyl oxidase, a gene whose product catalyses the deposition of collagen was also up-regulated by IL1alpha treatment (P<0.05), as was 11beta hydroxysteroid dehydrogenase type 1(11betaHSDt1). Genes down-regulated by IL1alpha included 3beta hydroxysteroid dehydrogenase and members of the alcohol dehydrogenase family (P<0.05). A panel of genes highly expressed in OSE cells under basal conditions were identified, including genes associated with multi-drug resistance in tumours, such as P-glycoprotein-1.
Normal OSE cells can modulate the inflammatory environment associated with ovulation. Genes involved include those required to facilitate repair of the ovarian surface as well as 11betaHSDt1, which can generate anti-inflammatory glucocorticoid as a means of countering inflammation. Additionally, patterns of gene expression under both basal and cytokine stimulated conditions have illuminated putative OSE functions, and identified a number of genes with implications for ovarian cancer studies (Supported by the Medical Research Council (Prg. Grant 0000066) and European Commission).
22 - 24 Mar 2004
British Endocrine Societies