title: Cationic lipid:DNA complexes allow bleomycin uptake by melanoma cells.
abstract: Bleomycin is a chemotherapeutic agent barely diffusible through the plasmatic membrane. We evaluated DNA/cationic lipids complexes (lipoplexes) as mediators of its uptake in four spontaneous canine melanoma derived cell lines (Ak, Bk, Br and Rkb). Cell survival after lipofection plus or minus bleomycin was determined by the acid phosphatase method and the cellular uptake of lipoplexes, carrying the E. coli β-galactosidase gene, was evidenced by SYBR Green I staining. The four cell lines resulted sensitive to the bleomycin/lipoplexes system in both spatial configurations. Survival rates values were lower than 20% in monolayers of the four tested lines and lower than 30% in three lines (Ak, Bk and Rkb) when grown as spheroids. The sensitization to bleomycin depended on lipoplexes in Ak and Rkb while Bk (in both spatial configurations) and Br (as monolayers) were sensitive to bleomycin alone. Although some degree of sensitivity to bleomycin was induced by cationic lipids alone in Ak and Rkb monolayers, the maximal bleomycin effects appeared in the presence of lipoplexes. The sensitization was independent of transcriptional activity. The co-administration of lipoplexes diminished bleomycin IC50: 10-fold in Ak and Rkb monolayers; and sensitized the Ak and Rkb resistant spheroids. The bleomycin cytotoxic effects depended on lipoplexes concentration and diminished when cells were incubated at 8°C. Our results suggest that lipoplexes sensitize cells to bleomycin, increasing its uptake by an active transport mechanism, such as endocytosis. The bleomycin/lipoplexes system appears as a promising combination of chemotherapy and non-viral cancer gene therapy.
authors: Unidad de Transferencia Genética, Instituto de Oncología Ángel H. Roffo, Universidad de Buenos Aires, Avenida San Martin 5481, 1417 Buenos Aires, Argentina.
authors: María L
authors: ML
authors: Gil-Cardeza
authors: Úrsula A
authors: ÚA
authors: Rossi
authors: Marcela S
authors: MS
authors: Villaverde
authors: Gerardo C
authors: GC
authors: Glikin
authors: Liliana M E
authors: LM
authors: Finocchiaro
mesh_terms: D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000494:pharmacology; D001693:Biological Transport, Active; D001761:Bleomycin / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000494:pharmacology; D002412:Cations; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000008:administration & dosage*; D004283:Dog Diseases / Q000188:drug therapy / Q000473:pathology; D004285:Dogs; D019008:Drug Resistance, Neoplasm; D004705:Endocytosis; D015316:Genetic Therapy / Q000379:methods; D020128:Inhibitory Concentration 50; D008055:Lipids / Q000737:chemistry; D008081:Liposomes; D008545:Melanoma / Q000188:drug therapy* / Q000473:pathology / Q000662:veterinary; D013696:Temperature
chemical_list: D000903:Antibiotics, Antineoplastic; D002412:Cations; D008055:Lipids; D008081:Liposomes; D001761:Bleomycin; D004247:DNA