Bcl-2 antagonizes apoptotic cell death induced by two new ceramide analogues.
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 > or = 14 microM 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.
Wieder
T
T
Department of Dermatology, University Medical Center Benjamin Franklin, The Free University of Berlin, Germany.
Geilen
C C
CC
Kolter
T
T
Sadeghlar
F
F
Sandhoff
K
K
Brossmer
R
R
Ihrig
P
P
Perry
D
D
Orfanos
C E
CE
Hannun
Y A
YA
D000577:Amides / Q000138:chemical synthesis / Q000494:pharmacology*; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D002518:Ceramides / Q000032:analysis / Q000138:chemical synthesis / Q000494:pharmacology*; D005047:Etoposide / Q000494:pharmacology; D019254:Genes, bcl-2; D006801:Humans; D015394:Molecular Structure; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D014162:Transfection; D014343:Trypan Blue / Q000378:metabolism; D014407:Tumor Cells, Cultured
C107960:4-(dodecanoylamino)decan-5-ol; D000577:Amides; D000972:Antineoplastic Agents, Phytogenic; D002518:Ceramides; C064769:N-acetylsphingosine; C107959:N-thioacetylsphingosine; D005047:Etoposide; D011065:Poly(ADP-ribose) Polymerases; D014343:Trypan Blue; D013110:Sphingosine