Volume 69, Issue 4, Pages 601-609 (April 2016) The UGT2B28 Sex-steroid Inactivation Pathway Is a Regulator of Steroidogenesis and Modifies the Risk of Prostate Cancer Progression  Anaïs Belledant, Hélène Hovington, Luciana Garcia, Patrick Caron, Hervé Brisson, Lyne Villeneuve, David Simonyan, Bernard Têtu, Yves Fradet, Louis Lacombe, Chantal Guillemette, Eric Lévesque  European Urology  Volume 69, Issue 4, Pages 601-609 (April 2016) DOI: 10.1016/j.eururo.2015.06.054 Copyright © 2015 European Association of Urology Terms and Conditions

Fig. 1 Simplified representation of the UGT2B28 enzymatic activity profile toward steroid hormones. ADT=androsterone; DHT=dihydrotestosterone; 3α-diol=androstane-3α, 17β-diol; G=glucuronide. European Urology 2016 69, 601-609DOI: (10.1016/j.eururo.2015.06.054) Copyright © 2015 European Association of Urology Terms and Conditions

Fig. 2 UGT2B28 localizes differently in prostate tumors compared with normal glands. (A) Immunohistochemistry using UGT2B28 antibody in normal prostate; (B) in prostatic intraepithelial neoplasia; (C) negative control, (D) in tumor of UGT2B28 (−/−) patients and in tumoral glands of UGT2B28 (+/+) patients with (E) nuclear and (F) cytoplasmic staining. In normal prostate, UGT2B28 is strongly expressed in the nucleus of basal cells (bold arrows) and shows moderate to low expression in the nucleus of secretory cells (thin arrows). Blue arrows indicate nuclei of secretory cells in which UGT2B28 was not detected. European Urology 2016 69, 601-609DOI: (10.1016/j.eururo.2015.06.054) Copyright © 2015 European Association of Urology Terms and Conditions

Fig. 3 Associations between UGT2B28 staining, UGT2B28 copy-number variations, and prostate cancer (PCa) prognostic factors. Tissue microarrays (TMAs) consisting of tumor samples from 239 individuals with PCa were analyzed with the UGT2B28 (Ab2321) polyclonal antibody. Association of UGT2B28 expression levels with (A) UGT2B28 (+/+) (no deletions) versus UGT2B28 (+/−) (1 deletion) in PCa patients, (B) prostate-specific antigen (PSA) levels at diagnosis, (C) tumor volume, (D) the presence of a positive margin, (E) nodal status, and (F) Gleason score. Correlation between UGT2B28 messenger RNA expression and Gleason score in (G) Taylor et al [22] and (H) The Cancer Genome Atlas data sets. The data are presented as the mean plus or minus standard error of the mean. Comparisons between experimental groups were performed by a two-tailed Student t test, a Kruskal-Wallis test, or Tukey-Kramer adjustment. *p ≤ 0.05; **p ≤ 0.01 mRNA=messenger RNA; PCa=prostate cancer; PSA=prostate-specific antigen; TCGA=The Cancer Genome Atlas. European Urology 2016 69, 601-609DOI: (10.1016/j.eururo.2015.06.054) Copyright © 2015 European Association of Urology Terms and Conditions

Fig. 4 (A) The influence of UGT2B28 germline status and tumor overexpression in prostate cancer patients. (B) In patients with an active or partially active UGT2B28 pathway, germline status [15] and tumor overexpression influence disease phenotype. In patients with a completely inactive UGT2B28 steroid-inactivation pathway, potent androgens and their metabolites are significantly reduced, whereas the adrenal precursor androstenedione is increased. BCR=biochemical recurrence; PSA=prostate-specific antigen. European Urology 2016 69, 601-609DOI: (10.1016/j.eururo.2015.06.054) Copyright © 2015 European Association of Urology Terms and Conditions