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

16 alpha-Hydroxy-dehydroepiandrosterone sulfate (16 alpha-OH-DHEAS) mainly originates from the fetus and serves as precursor for placental estriol biosynthesis. For conversion of 16 alpha-OH-DHEAS to estriol several intracellular enzymes are required. However, prior to enzymatic conversion, 16 alpha-OH-DHEAS must enter the cells by carrier mediated transport. To identify these carriers, uptake of 16 alpha-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 16 alpha-OH-DHEAS with an apparent K-m of 23.1 +/- 5.1 mu M and V-max of 485.0 +/- 39.1 pmol/mg protein/min, while stably transfected SOAT- and NTCP-HEK293 cells showed uptake only under sodium conditions with K-m of 319.0 +/- 59.5 mu M and V-max of 1465.8 +/- 118.8 pmol/mg protein/min for SOAT and K-m of 51.4 +/- 9.9 mu M and V-max of 1423.3 +/- 109.6 pmol/mg protein/min for NTCP. In contrast, stably transfected OATP2B1-HEK293 cells did not transport 16 alpha-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 16 alpha-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. (C) 2014 Elsevier Ltd. All rights reserved.