2. Academic Validation
  3. Transport of the placental estriol precursor 16α-hydroxy-dehydroepiandrosterone sulfate (16α-OH-DHEAS) by stably transfected OAT4-, SOAT-, and NTCP-HEK293 cells

Transport of the placental estriol precursor 16α-hydroxy-dehydroepiandrosterone sulfate (16α-OH-DHEAS) by stably transfected OAT4-, SOAT-, and NTCP-HEK293 cells

  • J Steroid Biochem Mol Biol. 2014 Sep;143:259-65. doi: 10.1016/j.jsbmb.2014.03.013.
H Schweigmann 1,  A Sánchez-Guijo 2,  B Ugele 3,  K Hartmann 4,  M F Hartmann 2,  M Bergmann 4,  C Pfarrer 5,  B Döring 1,  S A Wudy 2,  E Petzinger 1,  J Geyer 1,  G Grosser 6
Affiliations

Affiliations

  • 1 Institute of Pharmacology and Toxicology, Justus Liebig University of Giessen, 35392 Giessen, Germany.
  • 2 Steroid Research & Mass Spectrometry Unit, Division of Pediatric Endocrinology and Diabetology, Center of Child and Adolescent Medicine, Justus Liebig University of Giessen, 35392 Giessen, Germany.
  • 3 University Hospital, Ludwig Maximilians University of Munich, 80337 Munich, Germany.
  • 4 Institute of Veterinary Anatomy, Histology and Embryology, Justus Liebig University of Giessen, 35392 Giessen, Germany.
  • 5 Department of Anatomy, University of Veterinary Medicine, 30173 Hannover, Germany.
  • 6 Institute of Pharmacology and Toxicology, Justus Liebig University of Giessen, 35392 Giessen, Germany. Electronic address: [email protected].
PMID: 24717977 DOI: 10.1016/j.jsbmb.2014.03.013
Abstract

16α-Hydroxy-dehydroepiandrosterone sulfate (16α-OH-DHEAS) mainly originates from the fetus and serves as precursor for placental estriol biosynthesis. For conversion of 16α-OH-DHEAS to estriol several intracellular enzymes are required. However, prior to enzymatic conversion, 16α-OH-DHEAS must enter the cells by carrier mediated transport. To identify these carriers, uptake of 16α-OH-DHEAS by the candidate carriers organic anion transporter OAT4, sodium-dependent organic anion transporter SOAT, Na(+)-taurocholate cotransporting polypeptide NTCP, and organic anion transporting polypeptide OATP2B1 was measured in stably transfected HEK293 cells by LC-MS-MS. Furthermore, the study aimed to localize SOAT in the human placenta. Stably transfected OAT4-HEK293 cells revealed a partly sodium-dependent transport for 16α-OH-DHEAS with an apparent Km of 23.1 ± 5.1 μM and Vmax of 485.0 ± 39.1 pmol/mg protein/min, while stably transfected SOAT- and NTCP-HEK293 cells showed uptake only under sodium conditions with Km of 319.0 ± 59.5 μM and Vmax of 1465.8 ± 118.8 pmol/mg protein/min for SOAT and Km of 51.4 ± 9.9 μM and Vmax of 1423.3 ± 109.6 pmol/mg protein/min for NTCP. In contrast, stably transfected OATP2B1-HEK293 cells did not transport 16α-OH-DHEAS at all. Immunohistochemical studies and in situ hybridization of formalin fixed and paraffin embedded sections of human late term placenta showed expression of SOAT in syncytiotrophoblasts, predominantly at the apical membrane as well as in the vessel endothelium. In conclusion, OAT4, SOAT, and NTCP were identified as carriers for the estriol precursor 16α-OH-DHEAS. At least SOAT and OAT4 seem to play a functional role for the placental estriol synthesis as both are expressed in the syncytiotrophoblast of human placenta.

Keywords

DHEAS; OAT4; Placenta; SOAT; Steroid sulfate; Transport.

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