A human small cell lung carcinoma cell line, resistant to 4'-(9-acridinylamino)-methanesulfon-m-anisidide and cross-resistant to camptothecin with a high level of topoisomerase I.
N417/AMSA cells, about 80-fold resistant to mAMSA [4'-(9-acridinylamino)-methanesulfon-m-anisidide], were obtained by serial passages of the parental human small cell lung carcinoma NCI-N417 (N417/p) in stepwise drug concentrations. The N417/AMSA cells were found to be 114-, 100-, and 9-fold cross-resistant to the topoisomerase II (Topo II) inhibitors VM26, VP16 and Doxorubicin (DXR); they showed a 2-fold decrease in Topo II activity. Interestingly, N417/AMSA cells which exhibited a 3-fold increase in topoisomerase I (Topo I) activity were 28-fold cross-resistant to camptothecin (CPT), a specific inhibitor of Topo I. In order to investigate the cellular mechanisms leading to the development of resistance, the effects of mAMSA and CPT on parental and resistant cell lines were analysed by alkaline elution. A decrease in DNA single-strand breaks (DNA-SSB) was observed in N417/AMSA cells treated with mAMSA or CPT compared to parental cells. Similar differences were obtained in isolated nuclei, suggesting that no modification of mAMSA and CPT accumulation occurred in resistant cells. Topo I was purified from N417/p (Topo I/p) and N417/AMSA (Topo I/AMSA) cells in the exponential phase of growth, and the inhibitory effects of CPT on relaxation activities were determined. Topo I/AMSA was found to be about 7-fold less sensitive to CPT than Topo I/p, suggesting the possible involvement of a mutation outside the gene region sequenced (codons 420 to 642) or post-translational modifications of the Topo I enzyme. These data indicate that increased Topo I activity cannot be related to CPT resistance, and suggest that mAMSA can generate multiple cellular modifications which may be involved in resistance to various drugs.
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Laboratory of Clinical and Molecular Pharmacology, Institut Gustave Roussy, Villejuif, France.
Riou
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D000677:Amsacrine / Q000494:pharmacology*; D001483:Base Sequence; D002166:Camptothecin / Q000494:pharmacology*; D018288:Carcinoma, Small Cell / Q000201:enzymology / Q000473:pathology*; D017931:DNA Primers; D004264:DNA Topoisomerases, Type I / Q000235:genetics / Q000378:metabolism*; D004250:DNA Topoisomerases, Type II / Q000235:genetics / Q000378:metabolism; D004351:Drug Resistance; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008175:Lung Neoplasms / Q000201:enzymology / Q000473:pathology*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000378:metabolism; D017422:Sequence Analysis, DNA; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D012333:RNA, Messenger; D059004:Topoisomerase I Inhibitors; D059005:Topoisomerase II Inhibitors; D000677:Amsacrine; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II; D002166:Camptothecin