title: Aberrant expression and regulation of NR2F2 and CTNNB1 in uterine fibroids.
abstract: Uterine fibroids are the most common benign tumour afflicting women of reproductive age. Despite the large healthcare burden caused by fibroids, there is only limited understanding of the molecular mechanisms that drive fibroid pathophysiology. Although a large number of genes are differentially expressed in fibroids compared with myometrium, it is likely that most of these differences are a consequence of the fibroid presence and are not causal. The aim of this study was to investigate the expression and regulation of NR2F2 and CTNNB1 based on their potential causal role in uterine fibroid pathophysiology. We used real-time quantitative RT-PCR, western blotting and immunohistochemistry to describe the expression of NR2F2 and CTNNB1 in matched human uterine fibroid and myometrial tissues. Primary myometrial and fibroid smooth muscle cell cultures were treated with progesterone and/or retinoic acid (RA) and sonic hedgehog (SHH) conditioned media to investigate regulatory pathways for these proteins. We showed that NR2F2 and CTNNB1 are aberrantly expressed in fibroid tissue compared with matched myometrium, with strong blood vessel-specific localisation. Although the SHH pathway was shown to be active in myometrial and fibroid primary cultures, it did not regulate NR2F2 or CTNNB1 mRNA expression. However, progesterone and RA combined regulated NR2F2 mRNA, but not CTNNB1, in myometrial but not fibroid primary cultures. In conclusion, we demonstrate aberrant expression and regulation of NR2F2 and CTNNB1 in uterine fibroids compared with normal myometrium, consistent with the hypothesis that these factors may play a causal role uterine fibroid development.
authors.affiliation: Department of Obstetrics and Gynaecology, Royal Women's Hospital, University of Melbourne, Level 7, 20 Flemington Road, Parkville, Victoria 3052, Australia.
authors.forename: Marina
authors.initials: M
authors.lastname: Zaitseva
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Holdsworth-Carson
authors.forename: Luke
authors.initials: L
authors.lastname: Waldrip
authors.forename: Julia
authors.initials: J
authors.lastname: Nevzorova
authors.forename: Luciano
authors.initials: L
authors.lastname: Martelotto
authors.forename: Beverley J
authors.initials: BJ
authors.lastname: Vollenhoven
authors.forename: Peter A W
authors.initials: PA
authors.lastname: Rogers
mesh_terms: D000328:Adult; D001808:Blood Vessels / Q000378:metabolism / Q000473:pathology; D051837:COUP Transcription Factor II / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D005260:Female; D005498:Follicular Phase / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D053823:Hedgehog Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D018231:Leiomyomatosis / Q000098:blood supply / Q000378:metabolism* / Q000473:pathology / Q000601:surgery; D008183:Luteal Phase / Q000378:metabolism; D008875:Middle Aged; D009215:Myometrium / Q000098:blood supply / Q000378:metabolism* / Q000473:pathology; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D011374:Progesterone / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D014212:Tretinoin / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000098:blood supply / Q000378:metabolism* / Q000473:pathology; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism*
chemical_list: D051837:COUP Transcription Factor II; D053823:Hedgehog Proteins; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D011374:Progesterone; D011994:Recombinant Proteins; D014212:Tretinoin; D051176:beta Catenin; C495270:CTNNB1 protein, human; C509239:SHH protein, human; C095569:NR2F2 protein, human