title: A comparative study of the androgenic properties of progesterone and the progestins, medroxyprogesterone acetate (MPA) and norethisterone acetate (NET-A).
abstract: The importance of investigating the molecular mechanism of action of medroxyprogesterone acetate (MPA) and norethisterone acetate (NET-A), two clinically important progestins used in hormone therapy (HT), has been highlighted by clinical evidence showing that MPA and norethisterone (NET) increase the risk of the development of breast cancer in HRT users, and that MPA may increase susceptibility to- and transmission of HIV-1. The aim of this study was to compare the molecular mechanisms of action of MPA, NET-A and progesterone (Prog) via the androgen receptor (AR) in a cell line model that can minimize confounding factors such as the presence of other steroid receptors. This study is the first to determine accurate apparent Ki values for Prog, MPA and NET-A toward the human AR in COS-1 cells. The results reveal that these ligands have a similar binding affinity for the AR to that of the natural androgen 5α-dihydrotestosterone (DHT) (Ki's for DHT, Prog, MPA and NET-A are 29.4, 36.6, 19.4 and 21.9 nM, respectively). Moreover, in both transactivation and transrepression transcriptional assays we demonstrate that, unlike Prog, MPA and NET-A are efficacious AR agonists, with activities comparable to DHT. One of the most novel findings of our study is that NET-A, like DHT, induces the ligand-dependent interaction between the NH2- and COOH-terminal domains (N/C-interaction) of the AR independent of promoter-context, while MPA does not induce the N/C interaction on a classical ARE and does so only weakly on an AR-selective ARE. This suggests that MPA and NET-A may exert differential promoter-specific actions via the AR in vivo. Consistent with this, molecular modeling suggests that MPA and NET-A induce subtle differences in the structure of the AR ligand binding domain. Taken together, the results from this study suggest that unlike Prog, both MPA and NET-A used in hormonal therapy are likely to compete with DHT and exert significant and promoter-specific off-target transcriptional effects via the AR, possibly contributing to some of the observed side-effects with the clinical use of MPA and NET-A.
authors.affiliation: Department of Biochemistry, University of Stellenbosch, Private Bag X1, Matieland 7602, South Africa. Electronic address: drho@sun.ac.za.
authors.forename: Donita J
authors.initials: DJ
authors.lastname: Africander
authors.affiliation: Department of Biochemistry, University of Stellenbosch, Private Bag X1, Matieland 7602, South Africa.
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Storbeck
authors.affiliation: Department of Molecular and Cell Biology, University of Cape Town, Private Bag X3, Rondebosch 7701, South Africa. Electronic address: Janet.Hapgood@uct.co.za.
authors.forename: Janet P
authors.initials: JP
authors.lastname: Hapgood
mesh_terms: D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D003280:Contraceptives, Oral, Synthetic / Q000494:pharmacology; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D017258:Medroxyprogesterone Acetate / Q000494:pharmacology*; D009640:Norethindrone / Q000031:analogs & derivatives* / Q000494:pharmacology; D000077563:Norethindrone Acetate; D011374:Progesterone / Q000494:pharmacology*; D011372:Progestins / Q000494:pharmacology*; D011944:Receptors, Androgen / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: C410798:AR protein, human; D000728:Androgens; D018931:Antineoplastic Agents, Hormonal; D003280:Contraceptives, Oral, Synthetic; D011372:Progestins; D011944:Receptors, Androgen; D011374:Progesterone; D000077563:Norethindrone Acetate; D017258:Medroxyprogesterone Acetate; D008156:Luciferases; D009640:Norethindrone
keywords: Androgen receptor; Binding; Injectable contraceptives; N/C-interaction; Progestins; Transcription