Bcl‐2 antagonizes apoptotic cell death induced by two new ceramide analogues

FEBS Letters

Sandhoff, Konrad¹  Hannun, Yusuf A³  Wieder, Thomas²  Brossmer, Reinhard⁴  Orfanos, Constantin E²  Ihrig, Petra⁴  Geilen, Christoph C²  Kolter, Thomas¹  Perry, David³  Sadeghlar, Farsaneh¹ 

[1] Institute of Organic Chemistry and Biochemistry, University of Bonn, Bonn, Germany;Department of Dermatology, University Medical Center Benjamin Franklin, The Free University of Berlin, Hindenburgdamm 30, D-12200 Berlin, Germany;Department of Medicine, University Medical Center, Duke University, Durham, NC, USA;Institute of Biochemistry, University of Heidelberg, Heidelberg, Germany

关键词: Apoptosis;    Bcl-2;    Sphingomyelin cycle;    Synthetic ceramide;    C2-CerO;    N-acetylsphingosine;    C2-CerS;    N-thio-acetylsphingosine;    DAG;    diacylglycerol;    DETAPAc;    diethylenetriaminepentaacetic acid;    ECL;    enhanced chemiluminescence;    EGTA;    [ethylenebis(oxyethylenenitrilo)]tetraacetic acid;    FCS;    fetal calf serum;    FS-5;    4-dodecanoylamino-decan-5-ol;    IC50;    half-inhibitory concentration;    LD50;    half-lethal dosis;    Molt 4;    human leukemia cell line;    PAGE;    polyacrylamide gel electrophoresis;    PARP;    poly-(ADP-ribose)polymerase;    PBS;    phosphate-buffered saline;    RPMI;    Roswell Park Memorial Institute;    SDS;    sodium dodecylsulfate;    SM;    sphingomyelin;    TNFα;    tumor necrosis factor α;    TLC;    thin layer chromatography;   

DOI  :  10.1016/S0014-5793(97)00717-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Ceramides which arise in part from the breakdown of sphingomyelin comprise a class of antiproliferative lipids and have been implicated in the regulation of programmed cell death better known as apoptosis. In the present study, two new synthetic ceramide analogues, N-thioacetylsphingosine and FS-5, were used in Molt 4 cells to induce cell death. Besides their cytotoxic effects at concentrations ≥14 μM the data obtained clearly show that both analogues induced apoptosis at concentrations below this critical concentration as assessed by trypan blue exclusion and cleavage of the death substrate poly-(ADP-ribose) polymerase (PARP). Additional experiments in bcl-2-transfected Molt 4 cells revealed that the apoptotic but not the lytic effects of the analogues were antagonized by the apoptosis inhibitor Bcl-2. Furthermore, neither N-thio-acetylsphingosine nor FS-5 induced PARP cleavage in bcl-2-transfected Molt 4 cells indicating that the induction of apoptotic cell death by cell permeable ceramides is not due to unspecific disturbance of the cell membrane.

【 授权许可】

Unknown