Nicotine transport in a human choriocarcinoma cell line (JAR).
Smoking is a major health problem in pregnancy resulting in intrauterine growth retardation and birth complications. Nicotine, a toxic component of cigarette smoke, interferes with amino acid transport in the placenta and stimulates catecholamine release resulting in uteroplacental vasoconstriction. Transplacental transport of nicotine may be an important determinant of placental and fetal exposure. Our aim was to determine the mechanism of nicotine transport in the human choriocarcinoma cell line, JAR, as a model for the placenta. JAR cells were subcultured in 12-well plates following trypsinization at a seeding density of 0.5 x 10(6) cells/well (1.3 x 10(5) cells/cm2). Uptake studies of [3H]nicotine were carried out in JAR cell monolayers on day 2 after plating. [3H]Nicotine uptake was saturable (Km 156 microM), sensitive to temperature, and inhibited by unlabeled nicotine and various organic cations including mecamylamine and quinidine, but not by guanidine, tetraethylammonium (TEA), or neurotransmitters. Counterflux of [3H]nicotine uptake was produced by unlabeled nicotine and mecamylamine but not by cotinine or acetylcholine, consistent with a carrier-mediated transport process. The uptake could be driven by an inside-negative membrane potential or by an outwardly directed pH gradient. This is the first demonstration of a carrier-mediated transport mechanism for nicotine in a human cell line. This transport mechanism may have implications to the disposition of nicotine in the human placenta.
Zevin
S
S
Division of Clinical Pharmacology and Experimental Therapeutics and Department of Biopharmaceutical Sciences, University of California-San Francisco, San Francisco, California 94143, USA.
Schaner
M E
ME
Giacomini
K M
KM
D001692:Biological Transport; D002822:Choriocarcinoma / Q000378:metabolism*; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009538:Nicotine / Q000493:pharmacokinetics*; D011247:Pregnancy; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000378:metabolism*
D009538:Nicotine