Characterization of glucose transport and glucose transporters in the human choriocarcinoma cell line, BeWo.
In this study we have characterized 2-deoxyglucose (2DG) transport and hexose transporter expression in the human choriocarcinoma cell line, BeWo. 2DG uptake in BeWo cells displayed saturable kinetics (V(max), 29+/-1.5 nmol/min/mg protein;K(m), 1.5+/-0.02 m m) and was significantly inhibited in the presence of 2-deoxyglucose, mannose and 3- O -methyl glucose (all at a competing concentration of 30 m m) by up to 97 per cent, but not by galactose or fructose. Glucose uptake was not Na(+)-dependent, but was inhibited by cytochalasin B (by approx 85 per cent) indicating that hexose uptake was mediated via a facilitative glucose transport mechanism. Northern and immunoblot analyses revealed that BeWo cells expressed GLUT1 and GLUT5, but not GLUT2 or GLUT3. On immunoblots, GLUT1 migrated as a broad protein band on SDS-gels (average M(r)of 55 kDa) and treatment with N -glycanase resulted in a significant shift in its electrophoretic mobility; the core protein migrating as a 40 kDa band indicating that the carrier was heavily glycosylated. GLUT5 was detected as a discrete 60 kDa band and like GLUT1, the observed immunoreactive signal was lost when using antiserum that had been pre-adsorbed with the antigenic peptide. Our findings indicate that BeWo cells express a facilitative glucose transport system with characteristics broadly similar to those reported in isolated human placental membrane vesicles and that they are likely to serve as a useful experimental system for studying the regulation of placental glucose transport and transporter expression.
Shah
S W
SW
Department of Anatomy and Physiology, University of Dundee, Dundee, DD1 4HN, Scotland.
Zhao
H
H
Low
S Y
SY
Mcardle
H J
HJ
Hundal
H S
HS
D001692:Biological Transport / Q000502:physiology; D002822:Choriocarcinoma / Q000378:metabolism*; D003847:Deoxyglucose / Q000378:metabolism; D005260:Female; D005947:Glucose / Q000493:pharmacokinetics*; D051272:Glucose Transporter Type 1; D051296:Glucose Transporter Type 5; D006801:Humans; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D010920:Placenta / Q000378:metabolism*; D011247:Pregnancy; D014407:Tumor Cells, Cultured
D051272:Glucose Transporter Type 1; D051296:Glucose Transporter Type 5; D009004:Monosaccharide Transport Proteins; C495677:SLC2A1 protein, human; D003847:Deoxyglucose; D005947:Glucose