Author/Authors :
Massimo Di Vito، نويسنده , , Luisa Lenti، نويسنده , , Arno Knijn، نويسنده , , Egidio Iorio، نويسنده , , Federica D’Agostino، نويسنده , , Agnese Molinari، نويسنده , , Annarica Calcabrini، نويسنده , , Annarita Stringaro، نويسنده , , Stefania Meschini ، نويسنده , , Giuseppe Arancia، نويسنده , , Argante Bozzi، نويسنده , , Roberto Strom، نويسنده , , Franca Podo، نويسنده ,
Abstract :
The presence of nuclear magnetic resonance (NMR)-visible mobile lipid (ML) domains in apoptotic lymphoblasts suggests alterations in neutral lipid metabolism and compartmentation during programmed cell death. The detection of similar ML signals in activated lymphocytes raises questions about common mechanisms of ML formation during apoptosis and upon lymphoblast stimulation. Structure and subcellular localization of ML domains were therefore investigated by NMR, fluorescence and electron microscopy in Jurkat T-lymphoblasts either induced to apoptosis (by anthracyclines or dexamethasone or by serum deprivation) or activated by phorbol myristate acetate (PMA) plus ionomycin. ML contents in drug-treated cells correlated linearly with apoptosis, irrespective of the specific inducer and cell cycle arrest phase (r=0.993, P<0.001). Similar ML levels were measured in drug-induced apoptotic cells (A≈30–40%) and in non-apoptotic PMA/ionomycin-treated lymphoblasts (72 h). Lower ML contents were instead formed in serum-deprived apoptotic cells, with respect to controls. Increases in ML signals were associated, in either apoptotic or activated cells, with the accumulation of cytoplasmic, osmophilic lipid bodies (diameter≤1.0 μm), surrounded by own membrane, possessing intramembrane particles. The results support the hypothesis that ML are formed in the cytoplasm of drug-induced apoptotic cells during an early, ‘biochemically active’ phase of programmed cell death.
Keywords :
apoptosis , Lymphoblast activation , Mobile lipid , 1H nuclear magnetic resonance , Anthracycline , dexamethasone