title: A novel polymorphism in a forkhead box A1 (FOXA1) binding site of the human UDP glucuronosyltransferase 2B17 gene modulates promoter activity and is associated with altered levels of circulating androstane-3α,17β-diol glucuronide.
abstract: UDP glucuronosyltransferase 2B17 is present in the prostate, where it catalyzes the addition of glucuronic acid to testosterone and dihydrotestosterone and their metabolites androsterone and androstane-3α,17β-diol. Hence, changes in UGT2B17 gene expression may affect the capacity of the prostate to inactivate and eliminate male sex hormones. In this work, we identify a prevalent polymorphism, -155G/A, in the proximal promoter of the UGT2B17 gene. This polymorphism modulates UGT2B17 promoter activity, because luciferase-gene reporter constructs containing the -155A allele were 13-fold more active than those containing the -155G allele in prostate cancer LNCaP cells. The -155G/A polymorphism is contained within a putative binding site for the transcription factor Forkhead Box A1 (FOXA1). Using gene reporter, electromobility shift, and chromatin immunoprecipitation analyses, we show that FOXA1 binds to this site and stimulates the UGT2B17 promoter. Furthermore, down-regulation of FOXA1 in LNCaP cells substantially reduces UGT2B17 mRNA levels. The binding of FOXA1 and subsequent stimulation of the UGT2B17 promoter is greatly reduced in the presence of the -155G allele compared with the -155A allele. Consonant with its capacity to be stimulated by FOXA1, the UGT2B17 -155A allele, compared with the -155G allele, is associated with higher levels of circulating androstane-3α,17β-diol glucuronide. Although the initial phases of prostate cancer are androgen-dependent and UGT2B17 inactivates androgens, there was no association of the UGT2B17 -155G/A polymorphism with prostate cancer risk. In summary, this work identifies FOXA1 as an important regulator of UGT2B17 expression in prostate cancer LNCaP cells and identifies a polymorphism that alters this regulation.
authors.affiliation: Department of Clinical Pharmacology, Flinders University School of Medicine, Flinders Medical Centre, Bedford Park SA 5042, Australia.
authors.forename: Dong Gui
authors.initials: DG
authors.lastname: Hu
authors.forename: Dione
authors.initials: D
authors.lastname: Gardner-Stephen
authors.forename: Gianluca
authors.initials: G
authors.lastname: Severi
authors.forename: Philip A
authors.initials: PA
authors.lastname: Gregory
authors.forename: Joanna
authors.initials: J
authors.lastname: Treloar
authors.forename: Graham G
authors.initials: GG
authors.lastname: Giles
authors.forename: Dallas R
authors.initials: DR
authors.lastname: English
authors.forename: John L
authors.initials: JL
authors.lastname: Hopper
authors.forename: Wayne D
authors.initials: WD
authors.lastname: Tilley
authors.forename: Peter I
authors.initials: PI
authors.lastname: Mackenzie
mesh_terms: D015113:Androstane-3,17-diol / Q000031:analogs & derivatives* / Q000097:blood; D001665:Binding Sites / Q000502:physiology; D045744:Cell Line, Tumor; D014453:Glucuronosyltransferase / Q000235:genetics* / Q000378:metabolism; D051541:Hepatocyte Nuclear Factor 3-alpha / Q000097:blood / Q000235:genetics*; D006801:Humans; D008297:Male; D015778:Minor Histocompatibility Antigens; D011110:Polymorphism, Genetic / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011471:Prostatic Neoplasms / Q000097:blood / Q000235:genetics
chemical_list: C496499:FOXA1 protein, human; D051541:Hepatocyte Nuclear Factor 3-alpha; D015778:Minor Histocompatibility Antigens; D015113:Androstane-3,17-diol; C028895:androstane-3,17-diol glucuronide; D014453:Glucuronosyltransferase; C500513:UGT2B17 protein, human