title: Presence of organic anion transporters 3 (OAT3) and 4 (OAT4) in human adrenocortical cells.
abstract: Since the organic anion transporter-1 (OAT1) has been implicated in cortisol release from bovine and rat adrenal zona fasciculata cells, we addressed the question of whether OATs are present in human adrenal cortical cells. In the human adrenal cell line NCI-H295R, 24-h cortisol secretion increased up to 30-fold on exposure to forskolin. Incubation of forskolin-treated cells for 24 h with the OAT substrates probenecid, p-aminohippurate (PAH), glutarate or cimetidine inhibited cortisol release partly. RT-PCR did not reveal expression of human OAT1 and OAT2, but OAT3 and OAT4 mRNAs were detected in both NCI-H295R cells and human adrenal tissue. When human OAT3 (hOAT3) and hOAT4 were expressed in Xenopus laevis oocytes, only hOAT3 showed [3H]cortisol uptake in excess of that of water-injected control oocytes. Cortisol uptake via OAT3 was saturable with an apparent Kt of 2.4 microM. In NCI-H295R cells, [3H]estrone sulphate uptake was saturable, cis-inhibited by OAT substrates and trans-stimulated by preloading with glutarate or cortisol. Likewise, [3H]PAH uptake was cis-inhibited by estrone sulphate and trans-stimulated by preloading the cells with PAH, glutarate or cortisol, indicating functional expression of OATs in the plasma membrane of NCI-H295R cells.
authors.affiliation: Abt. Nephrologie und Rheumatologie, Georg-August-Universität Göttingen, Göttingen, Germany.
authors.forename: Abdul R
authors.initials: AR
authors.lastname: Asif
authors.forename: Jürgen
authors.initials: J
authors.lastname: Steffgen
authors.forename: Maria
authors.initials: M
authors.lastname: Metten
authors.forename: R Willi
authors.initials: RW
authors.lastname: Grunewald
authors.forename: Gerhard A
authors.initials: GA
authors.lastname: Müller
authors.forename: Andrew
authors.initials: A
authors.lastname: Bahn
authors.forename: Gerhard
authors.initials: G
authors.lastname: Burckhardt
authors.forename: Yohannes
authors.initials: Y
authors.lastname: Hagos
mesh_terms: D000302:Adrenal Cortex / Q000166:cytology / Q000378:metabolism*; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D005576:Colforsin / Q000494:pharmacology; D004970:Estrone / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D006801:Humans; D006854:Hydrocortisone / Q000493:pharmacokinetics; D009865:Oocytes / Q000378:metabolism; D029362:Organic Anion Transporters, Sodium-Independent / Q000378:metabolism*; D014316:Tritium; D014407:Tumor Cells, Cultured; D014982:Xenopus laevis
chemical_list: D029362:Organic Anion Transporters, Sodium-Independent; C498886:SLC22A9 protein, human; C114338:organic anion transport protein 3; D014316:Tritium; D005576:Colforsin; D004970:Estrone; D000324:Adrenocorticotropic Hormone; C017296:estrone sulfate; D006854:Hydrocortisone