title: Progesterone receptor-NFκB complex formation is required for progesterone-induced NFκB nuclear translocation and binding onto the p53 promoter.
abstract: We previously demonstrated that progesterone (P4) up-regulates p53 expression in human umbilical venous endothelial cells (HUVECs) through P4 receptor (PR) activation of extranuclear signaling pathways. However, the involvement of nuclear PR in P4-increased p53 expression is still unclear. Here, the molecular mechanism underlying PR-regulated p53 expression in HUVECs was investigated. Treatment with P4 increased nuclear factor of κ light polypeptide gene enhancer in B-cells inhibitor, α phosphorylation (IκBα and nuclear factor-κB (NFκB) nuclear translocation. Interestingly, P4 also increased PR-A, but not PR-B, nuclear translocation in HUVECs. Immunoprecipitation assay illustrated that P4 increased the formation of PR-A-NFκB complex in both the cytosol and the nucleus of HUVEC. Chromatin immunoprecipitation assay showed an interaction between PR and the NFκB binding motif on the p53 promoter. Ablation of the NFκB binding motif in the p53 promoter completely abolished P4-increased p53 promoter activity. In the absence of P4, overexpression of NFκB did not increase NFκB nuclear translocation. In contrast, treatment of NFκB-overexpressing HUVECs with P4 for only 4 hours, which is much shorter than the time (21.5 h) required for P4-induced IκBα phosphorylation, increased NFκB nuclear translocation. Blockade of PR activity abolished this effect. Taken together, these results uncover a novel role of PR for P4-induced NFκB nuclear translocation and suggest that PR-A-NFκB complex formation is required for NFκB nuclear translocation and binding onto the p53 promoter in HUVECs. Our data indicate that both nuclear and extranuclear signaling pathways of PR are involved in P4-regulated p53 expression in HUVECs.
authors.affiliation: Department of Physiology (S.-P.H., W.-S.L.), School of Medicine, College of Medicine, Taipei Medical University, Taipei 110, Taiwan; Graduate Institute of Physiology (H.-C.Y.), College of Medicine, National Taiwan University, Taipei 100, Taiwan; Graduate Institute of Medical Sciences (C.-T.K., H.-C.W., L.-C.C., Y.-N.H., W.-S.L.), College of Medicine, Taipei Medical University, Taipei 110, Taiwan; and Cancer Research Center (W.-S.L.), Taipei Medical University Hospital, Taipei 110, Taiwan.
authors.forename: Sung-Po
authors.initials: SP
authors.lastname: Hsu
authors.forename: Ho-Ching
authors.initials: HC
authors.lastname: Yang
authors.forename: Chun-Ting
authors.initials: CT
authors.lastname: Kuo
authors.forename: Heng-Ching
authors.initials: HC
authors.lastname: Wen
authors.forename: Li-Ching
authors.initials: LC
authors.lastname: Chen
authors.forename: Yen-Nien
authors.initials: YN
authors.lastname: Huo
authors.forename: Wen-Sen
authors.initials: WS
authors.lastname: Lee
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology*; D002070:Butadienes / Q000494:pharmacology; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004963:Estrenes / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D005663:Furans / Q000494:pharmacology; D005786:Gene Expression Regulation; D006727:Hormone Antagonists / Q000494:pharmacology; D006801:Humans; D015735:Mifepristone / Q000494:pharmacology; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009570:Nitriles / Q000494:pharmacology; D011374:Progesterone / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011485:Protein Binding; D011980:Receptors, Progesterone / Q000378:metabolism*; D013450:Sulfones / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
chemical_list: C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D002070:Butadienes; D004791:Enzyme Inhibitors; D004963:Estrenes; D005419:Flavonoids; D005663:Furans; D006727:Hormone Antagonists; D016328:NF-kappa B; D009570:Nitriles; D011980:Receptors, Progesterone; D013450:Sulfones; D016159:Tumor Suppressor Protein p53; C113580:U 0126; C076196:Org 31710; D015735:Mifepristone; D011374:Progesterone; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one