The metabolism of 11β-hydroxyandrostenedione by steroidogenic enzymes yields metabolites contributing to the androgen pool in prostate cancer
Date
2018-03
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Stellenbosch : Stellenbosch University
Abstract
ENGLISH ABSTRACT: This study describes:
• The development and validation of three ultra-performance convergence chromatography
tandem mass spectrometry (UPC2-MS/MS) analytical methods which were applied in the
detection and quantification of C19 and C21 steroids, including C11-oxy C19 and C11-oxy C21
steroids;
• The investigation into the contribution of adrenal 11β-hydroxyandrostenedione (11OHA4)
and 11β-hydroxytestosterone (11OHT) to the pool of active androgens in the prostate, by
following androgen metabolism in normal epithelial prostate PNT2, benign prostatic
hyperplasia (BPH-1) and prostate cancer LNCaP, C4-2B and VCaP cell models;
Steroid profiles revealed 11β-hydroxysteroid dehydrogenase type 2 (11βHSD2) activity in all
the cell models, confirmed in the conversion of 11OHA4 to 11keto-androstenedione
(11KA4), with reductive 17β-hydroxysteroid dehydrogenase (17βHSD) enzymes metabolising
11KA4, ultimately yielding 11keto-testosterone (11KT).
• The in vitro investigation into the inactivation, reactivation, glucuronidation and sulfation of
11OHA4, 11OHT and their downstream metabolites;
In prostate cancer (PCa) cell models, the conjugation of 11KT and 11ketodihydrotestosterone (11KDHT) were hampered compared to testosterone (T) and
dihydrotestosterone (DHT), while the inactivation and reactivation of the C11-oxy C19
steroids were less efficient than the C19 steroids in BPH-1 cells.
• The in vivo steroid profiles in PCa, BPH and castration-resistant prostate cancer (CRPC)
tissue and plasma of healthy and PCa patients;
Analyses of the C19 and C11-oxy C19 steroids, together with glucuronide and sulfate
conjugates, showed increased unconjugated levels of 11KT and 11KDHT in plasma of PCa
patients compared to a healthy subject, and 11OHA4, 11KT and 11KDHT levels were
prominent in PCa tissue, while downstream inactive C11-oxy 3α-reduced metabolites were
identified in BPH and CRPC tissue.
AFRIKAANSE OPSOMMING: Hierdie studie beskryf die volgende: • Die ontwikkeling en validering van drie UPC2-MS/MS analitiese metodes wat vervolgens toegepas is in die skeiding en kwantifisering van steroïedmetaboliete. • Die ondersoek na 11OHA4 and 11OHT se bydrae tot die aktiewe androgeen poel in die prostaat; Androgeen metabolisme is in normale epiteel prostaat PNT2, BPH-1 en prostaat kanker LNCaP, C4-2B en VCaP selmodelle ondersoek. Die steroïdprofiele bevestig die teenwoordigheid van 11βHSD2 aktiwiteit in al die modelle in die omsetting van 11OHA4 na 11KA4, sowel as die aktiwiteit van 17βHSD wat vervolgens die omsetting van 11KA4 na 11KT gekataliseer het. • Die in vitro inaktivering, heraktivering en konjugering van 11OHA4 en 11OHT metaboliete; In prostaatkanker selmodelle was 11KT en 11KDHT konjugering oneffektief in vergelyking met T en DHT, en die inaktivering van die C11-oksie C19 steroïede was nie optimaal in vergelyking met die C19 steroïede in BPH-1 selmodelle nie. • Die in vivo steroïedprofiele in prostaatkankerweefsel, BPH weefsel en kastrasieweerstandige prostaatkankerweefsel en in plasma van gesonde normale individuë en prostaatkanker pasïente, asook die teenwoordigheid van ongekonjugeerde en gekonjugeerde steroïedmetaboliete in sirkulasie; Analiese toon dat ongekonjugeerde 11KT en 11KDHT hoër was in die plasma van pasïente met prostaatkanker; 11OHA4, 11KT en 11KDHT vlakke was abnormaal hoog in die prostaatkankerweefsel, terwyl onaktiewe C11-oksie 3α-gereduseerde metaboliete geïdentifiseer was in BPH weefsel en in kastrasie-weerstandige prostaatkankerweefsel.
AFRIKAANSE OPSOMMING: Hierdie studie beskryf die volgende: • Die ontwikkeling en validering van drie UPC2-MS/MS analitiese metodes wat vervolgens toegepas is in die skeiding en kwantifisering van steroïedmetaboliete. • Die ondersoek na 11OHA4 and 11OHT se bydrae tot die aktiewe androgeen poel in die prostaat; Androgeen metabolisme is in normale epiteel prostaat PNT2, BPH-1 en prostaat kanker LNCaP, C4-2B en VCaP selmodelle ondersoek. Die steroïdprofiele bevestig die teenwoordigheid van 11βHSD2 aktiwiteit in al die modelle in die omsetting van 11OHA4 na 11KA4, sowel as die aktiwiteit van 17βHSD wat vervolgens die omsetting van 11KA4 na 11KT gekataliseer het. • Die in vitro inaktivering, heraktivering en konjugering van 11OHA4 en 11OHT metaboliete; In prostaatkanker selmodelle was 11KT en 11KDHT konjugering oneffektief in vergelyking met T en DHT, en die inaktivering van die C11-oksie C19 steroïede was nie optimaal in vergelyking met die C19 steroïede in BPH-1 selmodelle nie. • Die in vivo steroïedprofiele in prostaatkankerweefsel, BPH weefsel en kastrasieweerstandige prostaatkankerweefsel en in plasma van gesonde normale individuë en prostaatkanker pasïente, asook die teenwoordigheid van ongekonjugeerde en gekonjugeerde steroïedmetaboliete in sirkulasie; Analiese toon dat ongekonjugeerde 11KT en 11KDHT hoër was in die plasma van pasïente met prostaatkanker; 11OHA4, 11KT en 11KDHT vlakke was abnormaal hoog in die prostaatkankerweefsel, terwyl onaktiewe C11-oksie 3α-gereduseerde metaboliete geïdentifiseer was in BPH weefsel en in kastrasie-weerstandige prostaatkankerweefsel.
Description
Thesis (PhD)--Stellenbosch University, 2018.
Keywords
Adrenal androgens, Prostate cancer, Mass spectrometry, Adrenal-linked endocrine diseases, UCTD