Decreased drug accumulation without increased drug efflux in a novel MRP-overexpressing multidrug-resistant cell line.
KB/7D cells represent a multidrug-resistant subclone of human nasopharyngeal carcinoma KB cells generated by continuous exposure to the topoisomerase II inhibitor VP-16 (etoposide). KB/7D cells also show cross-resistance to doxorubicin and vincristine. Phenotypic traits of the cell line include a 2-fold decrease in topoisomerase II levels and a decrease in the uptake of VP-16 without an increase in the rate of drug efflux or expression of P-glycoprotein, suggesting a novel mechanism associated with the uptake of anticancer drugs. This study demonstrated that the multidrug-resistance associated protein (MRP) is overexpressed in KB/7D cells, and that the loss of resistance in revertant cells correlates with the loss of MRP. The resistance to VP-16 and doxorubicin could be overcome, partially, and resistance to vincristine could be overcome completely, by the L-enantiomer of verapamil, but not by the D-enantiomer or by BIBW 22 (4-[N-(2-hydroxy-2-methyl-propyl)-ethanolamino]-2,7-bis[cis-2,6-++ +dimethylmorpholino)-6-phenylpteridin), an inhibitor of MDR-1. L-Verapamil was shown to be significantly more potent than D-verapamil in modulating the accumulation defect in KB/7D cells towards doxorubicin, as measured by flow cytometry and confocal microscopy, and towards VP-16, as measured by increases in protein-linked DNA strand breaks. This suggests that KB/7D cells are multidrug resistant due to decreases in topoisomerase II levels and the overexpression of MRP, that MRP leads to a decrease in drug accumulation, and that L-verapamil can modulate the MRP-associated accumulation defect and drug-resistance phenotype. This contrasts with previous studies that suggest that MRP causes multidrug resistance by exporting cytotoxic drugs out of the cell and that did not show modulation of MRP by verapamil.
Gaj
C L
CL
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06510-8066, USA.
Anyanwutaku
I
I
Chang
Y H
YH
Cheng
Y C
YC
D018528:ATP-Binding Cassette Transporters / Q000096:biosynthesis / Q000502:physiology*; D000970:Antineoplastic Agents / Q000494:pharmacology; D002460:Cell Line; D004317:Doxorubicin / Q000494:pharmacology; D018432:Drug Resistance, Multiple; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005047:Etoposide / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D007624:KB Cells; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D059005:Topoisomerase II Inhibitors; D014700:Verapamil / Q000494:pharmacology*; D014750:Vincristine / Q000494:pharmacology
D018528:ATP-Binding Cassette Transporters; D000970:Antineoplastic Agents; D004317:Doxorubicin; D004791:Enzyme Inhibitors; D005047:Etoposide; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D059005:Topoisomerase II Inhibitors; D014700:Verapamil; D014750:Vincristine