title: Progesterone acts via the nuclear glucocorticoid receptor to suppress IL-1β-induced COX-2 expression in human term myometrial cells.
abstract: Progesterone is widely used to prolong gestation in women at risk of preterm labour (PTL), and acts at least in part via the inhibition of inflammatory cytokine-induced prostaglandin synthesis. This study investigates the mechanisms responsible for this inhibition in human myometrial cells. We used reporter constructs to demonstrate that interleukin 1beta (IL-1β) inhibits progesterone driven PRE activation via p65 activation and that IL-1β reduced progesterone driven gene expression (FKBP5). Conversely, we found that the activity of a p65-driven NFκB reporter construct was reduced by overexpression of progesterone receptor B (PRB) alone and that this was enhanced by the addition of MPA and that both MPA and progesterone suppressed IL-1β-driven cyclo-oxygenase-2 (COX-2) expression. We found that over-expressed Halo-tagged PRB, but not PRA, bound to p65 and that in IL-1β-treated cells, with no overexpression of either PR or p65, activated p65 bound to PR. However, we found that the ability of MPA to repress IL-1β-driven COX-2 expression was not enhanced by overexpression of either PRB or PRA and that although the combined PR and GR antagonist Ru486 blocked the effects of progesterone and MPA, the specific PR antagonist, Org31710, did not, suggesting that progesterone and MPA act via GR and not PR. Knockdown using siRNA confirmed that both MPA and progesterone acted via GR and not PR or AR to repress IL-1β-driven COX-2 expression. We conclude that progesterone acts via GR to repress IL-1β-driven COX-2 activation and that although the interaction between p65 and PRB may be involved in the repression of progesterone driven gene expression it does not seem to be responsible for progesterone repression of IL-1β-induced COX-2 expression.
authors.affiliation: Chelsea and Westminster Hospital, London, United Kingdom.
authors.forename: Kaiyu
authors.initials: K
authors.lastname: Lei
authors.forename: Li
authors.initials: L
authors.lastname: Chen
authors.forename: Ektoras X
authors.initials: EX
authors.lastname: Georgiou
authors.forename: Suren R
authors.initials: SR
authors.lastname: Sooranna
authors.forename: Shirin
authors.initials: S
authors.lastname: Khanjani
authors.forename: Jan J
authors.initials: JJ
authors.lastname: Brosens
authors.forename: Phillip R
authors.initials: PR
authors.lastname: Bennett
authors.forename: Mark R
authors.initials: MR
authors.lastname: Johnson
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D004963:Estrenes / Q000494:pharmacology; D005260:Female; D005663:Furans / Q000494:pharmacology; D005786:Gene Expression Regulation; D015971:Gene Expression Regulation, Enzymologic; D020868:Gene Silencing; D017930:Genes, Reporter; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism*; D015735:Mifepristone / Q000494:pharmacology; D009215:Myometrium / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D011374:Progesterone / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D011980:Receptors, Progesterone / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism
chemical_list: D004963:Estrenes; D005663:Furans; D053583:Interleukin-1beta; D016328:NF-kappa B; D034741:RNA, Small Interfering; D011965:Receptors, Glucocorticoid; D011980:Receptors, Progesterone; D051996:Transcription Factor RelA; C076196:Org 31710; D015735:Mifepristone; D011374:Progesterone; D051546:Cyclooxygenase 2