Uptake of L-triiodothyronine sulphate by human choriocarcinoma cell line, JAr.
This study investigated uptake of triiodothyronine sulphate (T3S) and interactions between uptake of T3S and triiodothyronine (T3) using the human choriocarcinoma cell line (JAr) as a model of placental transport. Cells were incubated at 37 degrees C with 30 pM 125I-T3 for 2 min with unlabelled T3 (0-30 microM) or T3S (0-1 mM). Addition of an excess unlabelled T3 (30 microM) or T3S (1 mM) reduced the initial rate of 125I-T3 uptake by 69.3+/-3.6 per cent (P<0.0001) and 52.9+/-7.8 per cent (P<0.0001), respectively. The calculated Michaelis constant (Km) for T3 uptake was 0.378+/-0.133 microM (n = 3) with a corresponding maximum velocity (Vmax) of 15.4+/-6.9 pmol/min/mg protein. Uptake of 125I-T3 was inhibited in a dose-dependent way by the addition of unlabelled T3S (0-1 mM). The calculated inhibition constant (Ki) for the inhibition of 125I-T3 uptake by T3S was 121.8+/-35.2 microM (n = 6). Saturable uptake of 125I-T3S by JAr cells was negligible. The T3S preparation incubated with the cells contained about 0.1 per cent T3, sufficient to explain the apparent inhibition of 125I-T3 uptake by unlabelled T3S. These results suggest that, in contrast to T3 uptake in these cells, JAr cells do not have a saturable uptake mechanism for T3S, and that T3S does not interact with the T3 transporter in these cells.
Bernus
I
I
Division of Chemical Pathology, Queensland Health Pathology Service, Royal Brisbane Hospital Campus, Australia. bernusi@health.qld.gov.au
Mitchell
A M
AM
Manley
S W
SW
Mortimer
R H
RH
D001692:Biological Transport; D002822:Choriocarcinoma / Q000378:metabolism*; D005260:Female; D006801:Humans; D007457:Iodine Radioisotopes; D007700:Kinetics; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D014284:Triiodothyronine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000378:metabolism*
D007457:Iodine Radioisotopes; D014284:Triiodothyronine; C037673:triiodothyronine sulfate