Uptake of dehydroepiandrosterone-3-sulfate by isolated trophoblasts from human term placenta, JEG-3, BeWo, Jar, BHK cells, and BHK cells transfected with human sterylsulfatase-cDNA

• Published in: The Journal of steroid biochemistry and molecular biology Vol. 71; no. 5; pp. 203 - 211
• Main Authors: Ugele, Bernhard, Simon, Sabine
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 31.12.1999; Elsevier Science
• Subjects: Arylsulfatases - genetics; Arylsulfatases - metabolism; Biological and medical sciences; Biological Transport; Cell Line; Dehydroepiandrosterone Sulfate - metabolism; DNA, Complementary - genetics; Embryology: invertebrates and vertebrates. Teratology; Female; Fetal membranes; Fundamental and applied biological sciences. Psychology; General aspects. Development. Fetal membranes; Humans; Kinetics; Placenta - metabolism; Steryl-Sulfatase; Taurocholic Acid - metabolism; Transfection; Trophoblasts - metabolism; Tumor Cells, Cultured; taurocholate, N-(2-sulfoethyl)-5β-cholan-3α,7α, 12α-triol-24-amide; steryl-sulfate sulfohydrolase (EC 3.1.6.2); DHEA-S, dehydroepiandrosterone-3-sulfate; STS, sterylsulfatase; 3β-sulfoxy-androst-5-ene-17-one; Human; Trophoblaste; Fetal membrane; Enzyme; Esterases; In vitro; Uptake; Dehydroepiandrosterone sulfate; Cell line; Placenta; Metabolic precursor; Steryl-sulfatase; Hydrolases; Sulfuric ester hydrolases
• ISSN: 0960-0760; 1879-1220
• DOI: 10.1016/S0960-0760(99)00138-7

The human placenta lacks the enzyme 17α-hydroxylase/17-20-lyase, and is thus unable to convert cholesterol into estrogens. Therefore estrogen synthesis of trophoblast cells depends on the supply of precursors such as dehydroepiandrosterone-3-sulfate (DHEA-S) and 16α-hydroxy-dehydroepiandrosterone-3-sulfate by maternal and fetal blood. To investigate the cellular internalisation of these anionic hydrophilic precursors, the uptake of [ 3H]-/[ 35S]-DHEA-S and [ 3H]-taurocholate by isolated cytotrophoblasts, cells of choriocarcinoma cell lines (JEG-3, BeWo, Jar), BHK and BHK cells transfected with human sterylsulfatase-cDNA (BHK-STS cells) was studied. Furthermore, the activity of sterylsulfatase of these cells in suspension and in corresponding cell homogenate was measured. During the first 5 min of incubation with [ 3H]-DHEA-S or [ 35S]-DHEA-S, radioactivity of cytotrophoblasts increased significantly, while radioactivity of JEG-3, Jar, BHK and BHK-STS cells did not increase. Radioactivity of BeWo cells increased slightly. For all cell types, there was no significant difference for uptake of either substrate. During incubation with [ 3H]-taurocholate, radioactivity of cytotrophoblasts did not increase. Sterylsulfatase activity of cytotrophoblast homogenate was significantly lower than that of cytotrophoblast suspension. Sterylsulfatase activity of BHK-STS, JEG-3 or BeWo cell homogenate was significantly higher than that of the corresponding cell suspension. In BHK and Jar cells sterylsulfatase activity was not detectable. Cytotrophoblasts take up DHEA-S without prior hydrolysis. BHK, BHK-STS, JEG-3, and Jar cells do not take up and BeWo cells slowly take up DHEA-S. In cytotrophoblasts extracellular DHEA-S rapidly gains access to intracellular sterylsulfatase, while in choriocarcinoma and BHK-STS cells access of DHEA-S to sterylsulfatase is limited. Our results indicate, that uptake by cytotrophoblasts is mediated by a carrier which is not expressed in choriocarcinoma or BHK cells and which is different from the known taurocholate-transporting organic anion transporting polypetides.