Two-step formation of 1H NMR visible mobile lipids during apoptosis of paclitaxel-treated K562 cells.
Despite increasing evidence on the formation of 1H NMR-detectable mobile lipid (ML) domains in cells induced to programmed cell death by continuous exposure to anticancer drugs, the time course of ML generation during the apoptotic cascade has not yet been fully elucidated. The present study shows that ML formation occurs at two different stages of apoptosis induced in human erythroleukemia K562 cells by a brief (3 hr) exposure to paclitaxel (Taxol), an antitumour drug with a stabilising effect on microtubules, or to paclitaxel plus tyrphostin AG957, a selective inhibitor of the p210(BCR-ABL) tyrosine kinase activity. A first wave of ML generation was in fact detected in paclitaxel-treated cells at the onset of the effector phase (8-24hr after exposure to the drug), plateaued at 24-48 hr and was eventually followed by further ML accumulation during the degradative phase (48-72 hr). Addition of AG957 to paclitaxel shifted to the 3-8 hr interval in both the early ML production and the onset of apoptotic events, such as chromatin condensation, phosphatidylserine externalization, cytochrome c release and caspase-3 activation. A significant loss of mitochondrial membrane potential was almost concomitant with the second wave of ML accumulation, associated in both cell systems with the phase of terminal cell degeneration, likely connected to non-regulated degradation of cell lipid components.
Brisdelli
Fabrizia
F
Department of Biomedical Sciences and Technologies, University of L'Aquila, Via Vetoio, Coppito 2, Italy.
Iorio
Egidio
E
Knijn
Arno
A
Ferretti
Amalia
A
Marcheggiani
Donatella
D
Lenti
Luisa
L
Strom
Roberto
R
Podo
Franca
F
Bozzi
Argante
A
D017304:Annexin A5 / Q000032:analysis; D000970:Antineoplastic Agents / Q000633:toxicity; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects*; D003576:Electron Transport Complex IV / Q000378:metabolism; D006801:Humans; D006859:Hydrogen; D007425:Intracellular Membranes / Q000187:drug effects / Q000502:physiology; D020014:K562 Cells; D007700:Kinetics; D050356:Lipid Metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology; D017239:Paclitaxel / Q000633:toxicity*
D017304:Annexin A5; D000970:Antineoplastic Agents; D006859:Hydrogen; D003576:Electron Transport Complex IV; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D017239:Paclitaxel