An innovative LC-MS/MS-based method for determining CYP 17 and CYP 19 activity in the adipose tissue of pre- and postmenopausal and ovariectomized women using 13C-labeled steroid substrates

J Clin Endocrinol Metab. 2014 Apr;99(4):1339-47. doi: 10.1210/jc.2013-3715. Epub 2014 Jan 23.

Abstract

Context: Does adipose tissue produce steroid hormones like an endocrine organ?

Object: To clarify whether adipose tissue produces sex steroid hormone like an endocrine organ, we estimated several key steroid hormone levels, as well as CYP17 and CYP19 activity, in ovariectomized, pre- and postmenopausal women by liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Subjects and methods: The subjects were 19 premenopausal (n = 12), postmenopausal (n = 4), and ovariectomized women (n = 3) aged 27-68 years. Serum, visceral adipose and sc adipose samples were taken from these subjects and stored at -70°C. The levels of cortisol, cortisone, progesterone (Prog), androstenedione, dehydroepiandrosterone, estrone, estradiol (E2), and T in serum and adipose tissue were estimated simultaneously by LC-MS/MS. CYP17 and CYP19 activity in tissues were assayed with the use of (13)C-labeled steroid precursors and LC-MS/MS-based estimation of the metabolites.

Results: E2 and Prog levels in the sera of postmenopausal or ovariectomized women were less than 10% of those in premenopausal women. No marked variations were seen in other hormones. Estrone, androstenedione, dehydroepiandrosterone, and Prog levels in the visceral and sc tissues of postmenopausal and ovariectomized women were 9-60 times higher than those in serum, whereas E2 and T levels were 3- to 7-fold higher than those in serum, and cortisol and cortisone levels were 20% of those found for serum. CYP17 in adipose tissue was found to have 17-hydroxylase and 20,17-lyase activity, with each catalytic activity being essentially equal. Therefore, CYP17 in adipose tissue is of the testicular/ovarian type but not adrenal type, which has 17-hydroxylase activity dominant. The presence of CYP19 activity in adipose tissue was approximately 3% of CYP17.

Conclusion: Our findings suggest that adipose tissue acts as an endocrine organ, with CYP17 and CYP19 activity playing an essential role in sex steroid hormone biosynthesis.

Publication types

  • Comparative Study

MeSH terms

  • Adipose Tissue / chemistry*
  • Adipose Tissue / enzymology
  • Adult
  • Aged
  • Aromatase / analysis*
  • Aromatase / metabolism
  • Carbon Isotopes / analysis
  • Chromatography, Liquid
  • Female
  • Humans
  • Middle Aged
  • Ovariectomy
  • Postmenopause* / metabolism
  • Premenopause* / metabolism
  • Steroid 17-alpha-Hydroxylase / analysis*
  • Steroid 17-alpha-Hydroxylase / metabolism
  • Tandem Mass Spectrometry / methods*

Substances

  • Carbon Isotopes
  • Aromatase
  • Steroid 17-alpha-Hydroxylase