Localization of adriamycin in model and natural membranes: influence of lipid molecular packing

• Published in: European journal of biochemistry Vol. 181; no. 3; pp. 695 - 702
• Main Authors: DUPOU-CEZANNE, L, SAUTEREAU, A.-M, TOCANNE, J.-F
• Format: Journal Article
• Language: English
• Published: Oxford Blackwell 15.05.1989
• Subjects: Animals; Antineoplastic agents; Biological and medical sciences; Cattle; Cell Line; Cell Membrane Permeability; Cricetinae; Cricetulus; Doxorubicin - metabolism; Fatty Acids - metabolism; Female; General aspects; Lipid Bilayers - metabolism; lipid membranes; Liposomes - metabolism; Medical sciences; Membrane Lipids - metabolism; Models, Theoretical; Ovary; Pharmacology. Drug treatments; Phosphatidylcholines - metabolism; Phosphatidylserines - metabolism; Phospholipids - metabolism; Spectrometry, Fluorescence - methods; Antineoplastic agent; Rodentia; Liposome; Phospholipid; Molecular interaction; Artificial membrane; CHO cell line; Fluorescence spectrometry; Vertebrata; Antibiotic; Mammalia; Luminescence quenching; Plasma membrane; Anthracyclins; Localization; Mechanism of action; Energy transfer; Hamster
• ISSN: 0014-2956; 1432-1033
• DOI: 10.1111/j.1432-1033.1989.tb14779.x

The interaction of adriamycin with lipids was studied in model (monolayers, small unilamellar vesicles, large multilamellar vesicles) and natural (chinese hamster ovary cell) membranes by measurement of fluorescence energy transfer and fluorescence quenching. 2-APam, 7-ASte, 12-ASte and anthracene-phosphatidylcholine were used as fluorescent probes in which the anthracene group is well located at graded depths in the membrane. Egg-yolk phosphatidylcholine and a 1/1 mixture of it with bovine brain phosphatidylserine were used in model membrane systems. Large fluorescence energy transfer was observed between these molecules as donors and the drug as acceptor. With liposomes, at pH 7.4 and over an adriamycin concentration range of 0-100 microM, the efficiency of energy transfer was 12-ASte greater than 7-ASte greater than 2-APam, with 100% energy transfer for 12-ASte above a drug concentration of 30 microM. At pH 5, where the fatty acids are buried deeper (0.45 nm) in the lipid bilayer due to protonation of the carboxyl group, the order of energy transfer 7-ASTe greater than 12-ASte = 2-APam was observed. Measurements of fluorescence quenching using the non-permeant Cu2+ ion as quencher and spectrophotometric assays indicated that around 40% of the adriamycin molecules were deeply embedded in the lipid bilayer. Adriamycin molecules thus appear to penetrate the lipid bilayer, with the aminoglycosyl group interacting with the lipid phosphate groups and the dihydroanthraquinone residue in contact with the lipid fatty acid chains. In contrast, fluorescence energy transfer and quenching studies on CHO cells showed that adriamycin penetrated the plasma membrane of these cells to a much more limited extent than in the model membrane systems. This can be related to the squeezing out of the drug from a film of phosphatidylcholine which was observed in monolayers by means of surface pressure, potential and fluorescence experiments. These observations indicated that the penetration of adriamycin into lipid bilayers strongly depends on the molecular packing of the lipid.