title: Anthracycline-induced DNA breaks and resealing in doxorubicin-resistant murine leukemic P388 cells.
abstract: Energy-dependent drug efflux is believed to be a major factor in cellular resistance to doxorubicin (DOX). However, recent studies have shown that decreased retention alone cannot account for anthracycline resistance, and possibly other factors, such as drug metabolism, free radical scavengers, and altered DNA damage/repair, may be involved. We have measured DOX-induced DNA damage and its repair in P388 cells sensitive (P388/S) and resistant (P388/R) to DOX. Our studies show 2- to 5-fold less DNA damage, measured as protein-associated single-strand DNA breaks, in P388/R cells when compared to similarly treated P388/S cells. The repair of DNA in whole cells, expressed as percent DNA rejoined, was complete in 4 hr in P388/R, whereas no repair was seen in P388/S cells until 20 hr. No difference in repair of DNA lesions was observed when nuclei were used in repair experiments. The absence of repair in sensitive whole cells may be due to high retention or slow drug efflux. Increase of cellular DOX retention by exposure of cells to trifluoperazine (TFP) or verapamil (VPL) did not result in the increase of DNA damage in P388/R cells. DOX analogs, N-trifluoroacetyladriamycin-14-valerate (AD 32), 4'-O-tetrahydropyranyladriamycin (THP-adriamycin), and N-benzyladriamycin-14-valerate (AD 198), induced 2- to 4-fold more DNA damage than DOX in resistant cells. There was no difference in the poly(ADP-ribose) synthesis of P388/S and P388/R cells exposed to DOX or AD 32. Since ADP-ribose polymer synthesis is associated with free radical-induced DNA damage and is indicative of DNA repair by an excision-repair mechanism, data from these studies suggest that DNA breaks in anthracycline-exposed cells may not be due to free radical production but rather to other mechanisms, such as inhibition of DNA topoisomerase II activity. The present studies, in addition to emphasizing the role of DNA damage in resistance, also underscore the relative importance of DNA topoisomerase II function in anthracycline cytotoxicity.
authors.affiliation: Division of Experimental Therapeutics, Papanicolaou Comprehensive Cancer Center, Miami, FL.
authors.forename: N
authors.initials: N
authors.lastname: Maniar
authors.forename: A
authors.initials: A
authors.lastname: Krishan
authors.forename: M
authors.initials: M
authors.lastname: Israel
authors.forename: T S
authors.initials: TS
authors.lastname: Samy
mesh_terms: D000818:Animals; D000903:Antibiotics, Antineoplastic; D002460:Cell Line; D002467:Cell Nucleus / Q000187:drug effects; D004249:DNA Damage; D004260:DNA Repair; D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D004317:Doxorubicin / Q000633:toxicity*; D004351:Drug Resistance; D066298:In Vitro Techniques; D007942:Leukemia, Experimental; D051379:Mice; D009279:Naphthacenes / Q000633:toxicity; D011064:Poly Adenosine Diphosphate Ribose / Q000096:biosynthesis; D013329:Structure-Activity Relationship
chemical_list: D000903:Antibiotics, Antineoplastic; D009279:Naphthacenes; D011064:Poly Adenosine Diphosphate Ribose; D004317:Doxorubicin; D004264:DNA Topoisomerases, Type I