title: Improved cellular accumulation is characteristic of anthracyclines which retain high activity in multidrug resistant cell lines, alone or in combination with verapamil or cyclosporin A.
abstract: We have examined the cellular accumulation of anthracycline compounds, alone or in conjunction with resistance modifiers, in an attempt to identify mechanisms by which multidrug resistance (MDR) can be circumvented. This was facilitated by using the EMT6 mouse mammary tumour cell line EMT6/P and its MDR subline EMT6/AR1.0 with 30-fold resistance to Adriamycin (ADM), and the human small cell lung cancer line H69/P together with its MDR subline H69/LX4 with 100-fold resistance to ADM. Both MDR lines hyperexpress membrane P-170 glycoprotein. The accumulation of ADM was compared to that seen for the anthracycline analogues aclacinomycin A (ACL), Ro 31-1215 and 4'-deoxy-4'-iodo-Adriamycin (iodo-ADM). These analogues were selected because of their high activity against MDR sublines, including H69/LX4 and EMT6/AR1.0. Both MDR cell lines exhibited a deficiency in ADM accumulation compared to the parent lines. Smaller differentials were seen using Ro 31-1215 or iodo-ADM. Both resistant sublines were able to accumulate ACL in identical amounts to their respective parental sublines. Improved drug accumulation is likely to contribute to the improved activity of the analogues against MDR cell lines. However, the relative accumulation defects in the resistant lines did not correlate exactly with the degree of resistance to a particular compound. Cyclosporin A (5 micrograms/ml) or verapamil (3.3 micrograms/ml) caused a preferential increase in uptake in both MDR sublines, with a small or negligible effect for the parental line. A smaller effect was observed with iodo-ADM and Ro 31-1215, and levels of ACL were unchanged in the MDR lines in the presence of either resistance modifier. These results indicate two mechanisms for circumventing drug resistance due to reduced drug accumulation. Structurally modified derivatives can partially or completely eliminate uptake differentials between parent and drug resistant cell lines. Any residual uptake can be eliminated using resistance modifiers. The two mechanisms may both operate via inhibition or circumvention of P-170 mediated efflux. The situation is complex, however, and this study indicates the possible involvement of additional resistance mechanisms.
authors.affiliation: MRC Clinical Oncology and Radiotherapeutics Unit, Cambridge, U.K.
authors.forename: H M
authors.initials: HM
authors.lastname: Coley
authors.forename: P R
authors.initials: PR
authors.lastname: Twentyman
authors.forename: P
authors.initials: P
authors.lastname: Workman
mesh_terms: D015250:Aclarubicin / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D003524:Cyclosporins / Q000378:metabolism*; D004317:Doxorubicin / Q000378:metabolism; D004338:Drug Combinations; D004351:Drug Resistance; D006801:Humans; D051379:Mice; D014407:Tumor Cells, Cultured / Q000378:metabolism; D014700:Verapamil / Q000378:metabolism*
chemical_list: D000903:Antibiotics, Antineoplastic; D003524:Cyclosporins; D004338:Drug Combinations; C011157:aclacinomycins; D015250:Aclarubicin; D004317:Doxorubicin; D014700:Verapamil