Progressive resistance to doxorubicin in mouse leukemia L1210 cells with multidrug resistance phenotype: reductions in drug-induced topoisomerase II-mediated DNA cleavage.
Cells selected for resistance to doxorubicin (DOX) express the multidrug resistance (MDR) phenotype, and resistance has been suggested to be due primarily to enhanced cellular efflux of drug. A progressively DOX-resistant (10- and 40-fold) L1210 mouse leukemia model system, which does not exhibit enhanced DOX efflux as a primary mechanism of resistance, was found to display the MDR phenotype, based on overexpression of P-glycoprotein in western blots and cross-resistance to vinca alkaloids. Cross-resistance to another topoisomerase II inhibitor, etoposide (VP-16), was similar to that of DOX (10- and 40-fold), whereas resistance to N-[4-(9-acridinylamino)-3-methoxyphenyl]methanesulfonamide (m-AMSA) was 5-fold lower. In contrast, no cross-resistance to camptothecin, an inhibitor of topoisomerase I, was observed. Topoisomerase II decatenation activity in nuclear extracts from 10- and 40-fold DOX-resistant cells was 2- and 4-fold lower, respectively, when compared to sensitive cells. In these cells, however, marked reductions in m-AMSA- and VP-16-induced topoisomerase II mediated DNA cleavage were found to exceed decreases in the catalytic activity of the enzyme. Results from this study demonstrated that, in progressively DOX-resistant L1210 mouse leukemia cells with the MDR phenotype, a better relation existed between the degree of resistance and reduced VP-16- and m-AMSA-induced topoisomerase II mediated DNA cleavage, than between increases in P-glycoprotein and concomitant reduction in DOX accumulation.
Ganapathi
R
R
Research Institute, Cleveland Clinic Foundation, OH 44195.
Grabowski
D
D
Ford
J
J
Heiss
C
C
Kerrigan
D
D
Pommier
Y
Y
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000677:Amsacrine / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology; D002470:Cell Survival / Q000187:drug effects; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism*; D004317:Doxorubicin / Q000494:pharmacology*; D004351:Drug Resistance / Q000235:genetics*; D005047:Etoposide / Q000494:pharmacology; D007939:Leukemia L1210 / Q000201:enzymology*; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000302:isolation & purification; D051379:Mice; D009363:Neoplasm Proteins / Q000096:biosynthesis; D010641:Phenotype; D013539:Simian virus 40; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D004279:DNA, Viral; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D000677:Amsacrine; D005047:Etoposide; D004317:Doxorubicin; D004250:DNA Topoisomerases, Type II