2-deoxy-D-glucose toxicity and transport in human multidrug-resistant KB carcinoma cell lines.
It is shown that a series of colchicine-selected multidrug-resistant (MDR) human KB carcinoma cell lines displayed increasing 2-deoxy-D-glucose collateral sensitivity, which correlated with increasing multidrug resistance. The relative resistance of MDR cell lines to 2-deoxy-D-glucose was reduced to 0.73 (KB-8-5), 0.3 (KB-8-5-11) and 0.2 (KB-C1) when compared with parental KB-3-1 (1.0). 2-Deoxy-D-glucose accumulation was found to be reduced in the MDR cell lines in a manner that correlated with 2-deoxy-D-glucose collateral sensitivity. At 30 min 2-deoxy-D-glucose accumulation was reduced to 0.61 (KB-8-5), 0.41 (KB-8-5-11) and 0.22 (KB-C1) relative to KB-3-1 uptake (1.0). The efflux of 2-deoxy-D-glucose was not significantly different between resistant and sensitive cell lines. Analysis of 2-deoxy-D-glucose uptake kinetics, by initial rate measurements, showed alterations in K(t) and J(max) for MDR when compared with KB-3-l cells. The levels of GLUT-1 facilitative transporter were found to be reduced significantly in the MDR cell lines in total cell homogenate and plasma membrane fractions by using Western blot analysis. Changes in the plasma membrane level of GLUT-1 correlated with 2-deoxy-D-glucose toxicity and uptake for MDR cell lines, where relative GLUT-i levels were reduced to 0.71 (KB-8-5), 0.43 (KB-8-5-1 1) and 0.27 (KB-Cl) relative to KB-31(1.0). It is concluded that the response of human KB MDR cells to 2-deoxy-D-glucose involved alterations in the level and activity of the facilitative glucose transporter, GLUT-1, in a manner that is associated with the degree of multidrug resistance.
Bentley
J
J
Department of Biology, The University of York, UK.
Bell
S E
SE
Quinn
D M
DM
Kellett
G L
GL
Warr
J R
JR
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000345:Affinity Labels / Q000378:metabolism; D001386:Azides / Q000378:metabolism; D001667:Binding, Competitive; D015153:Blotting, Western; D003078:Colchicine; D003847:Deoxyglucose / Q000493:pharmacokinetics* / Q000494:pharmacology; D004095:Dihydropyridines / Q000378:metabolism; D018432:Drug Resistance, Multiple; D019008:Drug Resistance, Neoplasm; D006801:Humans; D010693:Phloretin / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D014410:Tumor Stem Cell Assay
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000345:Affinity Labels; D001386:Azides; D004095:Dihydropyridines; C041130:azidopine; D003847:Deoxyglucose; D010693:Phloretin; D003078:Colchicine