Limited in vitro efficacy of CYP17A1 inhibition on human castration resistant prostate cancer

Takeo Kosaka, Akira Miyajima, Yota Yasumizu, Yasumasa Miyazaki, Eiji Kikuchi, Mototsugu Oya

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Although accumulating evidence indicates high expression of CYP17A1(P45017A1) allows castration resistant prostate cancer (CRPC) to maintain high intratumoral androgen levels, the potential P45017A1 activity has not been characterized yet. The aim of this study was to examine the potential CYP17A1 activity including 17α-hydroxylase and 17,20-lyase activities in human CRPC and the effect of a CYP17A inhibitor. We used three human CRPC cell lines: C4-2 and C4-2AT6 which was established from C4-2 under androgen ablation conditions for 6 months, and PC3. To ascertain the potential CYP17A1 activity, we cultured with the steroid precursors: 13C-[2,3,4]-progesterone (13C-Prog), and analyzed the sequential biosynthesis 13C-[2,3,4]-17-hydroxyprogesterone (13C-17OHP) and 13C-[2,3,4]-androstenedione(13C-Adione) by liquid chromatography/mass spectrometry (LC/MS/MS).The C4-2AT6 cells showed significantly higher CYP17A1 expression than C4-2 cells (p < 0.001). LC/MS/MS analysis enabled us to detect the 13C-17-OHP and 13C-A-dione in these cell lines. The concentration ratio of 13C-Adione/13C-17OHP (Adione-17OHP ratio), which is thought to reflect the differences between 17-hydroxylase and 17,20-lyase activities, was then determined. The Adione-17OHP ratio in C4-2AT6 cells was significantly higher than that of C4-2 cells (p < 0.001). Abiraterone were able to inhibit the CYP17A activities, although abiraterone did not have anti-proliferative effects on C4-2 and C4-2AT6 cells at clinically achievable concentrations of <1000 nM in vitro. The present study clearly demonstrates CRPC have the dual activities of CYP17A1 mediated by 17-hydroxylase activity and 17,20-lyase activity. Abiraterone doesn't have an in vitro anti-proliferative efficacy in CRPC cells, suggesting limited efficacy in vitro.

Original languageEnglish
Pages (from-to)39-44
Number of pages6
JournalSteroids
Volume92
DOIs
Publication statusPublished - 2014 Dec 15

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Steroid 17-alpha-Hydroxylase
Castration
Mixed Function Oxygenases
17-alpha-Hydroxyprogesterone
Prostatic Neoplasms
Androstenedione
Androgens
Cells
Biosynthesis
Liquid chromatography
Ablation
Mass spectrometry
Progesterone
Steroids
Cell Line
Human Activities
Liquid Chromatography
abiraterone
In Vitro Techniques
Mass Spectrometry

Keywords

  • Abiraterone
  • C4-2AT6
  • Castration-resistant prostate cancer
  • CYP17A1
  • Liquid chromatography/mass spectrometry
  • P45017A1

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Endocrinology
  • Molecular Biology
  • Organic Chemistry
  • Pharmacology

Cite this

Limited in vitro efficacy of CYP17A1 inhibition on human castration resistant prostate cancer. / Kosaka, Takeo; Miyajima, Akira; Yasumizu, Yota; Miyazaki, Yasumasa; Kikuchi, Eiji; Oya, Mototsugu.

In: Steroids, Vol. 92, 15.12.2014, p. 39-44.

Research output: Contribution to journalArticle

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abstract = "Although accumulating evidence indicates high expression of CYP17A1(P45017A1) allows castration resistant prostate cancer (CRPC) to maintain high intratumoral androgen levels, the potential P45017A1 activity has not been characterized yet. The aim of this study was to examine the potential CYP17A1 activity including 17α-hydroxylase and 17,20-lyase activities in human CRPC and the effect of a CYP17A inhibitor. We used three human CRPC cell lines: C4-2 and C4-2AT6 which was established from C4-2 under androgen ablation conditions for 6 months, and PC3. To ascertain the potential CYP17A1 activity, we cultured with the steroid precursors: 13C-[2,3,4]-progesterone (13C-Prog), and analyzed the sequential biosynthesis 13C-[2,3,4]-17-hydroxyprogesterone (13C-17OHP) and 13C-[2,3,4]-androstenedione(13C-Adione) by liquid chromatography/mass spectrometry (LC/MS/MS).The C4-2AT6 cells showed significantly higher CYP17A1 expression than C4-2 cells (p < 0.001). LC/MS/MS analysis enabled us to detect the 13C-17-OHP and 13C-A-dione in these cell lines. The concentration ratio of 13C-Adione/13C-17OHP (Adione-17OHP ratio), which is thought to reflect the differences between 17-hydroxylase and 17,20-lyase activities, was then determined. The Adione-17OHP ratio in C4-2AT6 cells was significantly higher than that of C4-2 cells (p < 0.001). Abiraterone were able to inhibit the CYP17A activities, although abiraterone did not have anti-proliferative effects on C4-2 and C4-2AT6 cells at clinically achievable concentrations of <1000 nM in vitro. The present study clearly demonstrates CRPC have the dual activities of CYP17A1 mediated by 17-hydroxylase activity and 17,20-lyase activity. Abiraterone doesn't have an in vitro anti-proliferative efficacy in CRPC cells, suggesting limited efficacy in vitro.",
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AU - Kosaka, Takeo

AU - Miyajima, Akira

AU - Yasumizu, Yota

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AU - Kikuchi, Eiji

AU - Oya, Mototsugu

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N2 - Although accumulating evidence indicates high expression of CYP17A1(P45017A1) allows castration resistant prostate cancer (CRPC) to maintain high intratumoral androgen levels, the potential P45017A1 activity has not been characterized yet. The aim of this study was to examine the potential CYP17A1 activity including 17α-hydroxylase and 17,20-lyase activities in human CRPC and the effect of a CYP17A inhibitor. We used three human CRPC cell lines: C4-2 and C4-2AT6 which was established from C4-2 under androgen ablation conditions for 6 months, and PC3. To ascertain the potential CYP17A1 activity, we cultured with the steroid precursors: 13C-[2,3,4]-progesterone (13C-Prog), and analyzed the sequential biosynthesis 13C-[2,3,4]-17-hydroxyprogesterone (13C-17OHP) and 13C-[2,3,4]-androstenedione(13C-Adione) by liquid chromatography/mass spectrometry (LC/MS/MS).The C4-2AT6 cells showed significantly higher CYP17A1 expression than C4-2 cells (p < 0.001). LC/MS/MS analysis enabled us to detect the 13C-17-OHP and 13C-A-dione in these cell lines. The concentration ratio of 13C-Adione/13C-17OHP (Adione-17OHP ratio), which is thought to reflect the differences between 17-hydroxylase and 17,20-lyase activities, was then determined. The Adione-17OHP ratio in C4-2AT6 cells was significantly higher than that of C4-2 cells (p < 0.001). Abiraterone were able to inhibit the CYP17A activities, although abiraterone did not have anti-proliferative effects on C4-2 and C4-2AT6 cells at clinically achievable concentrations of <1000 nM in vitro. The present study clearly demonstrates CRPC have the dual activities of CYP17A1 mediated by 17-hydroxylase activity and 17,20-lyase activity. Abiraterone doesn't have an in vitro anti-proliferative efficacy in CRPC cells, suggesting limited efficacy in vitro.

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