Dissociation of endogenous cellular ceramide from NF-kappa B activation.
The participation of cell ceramide in tumor necrosis factor (TNF)-alpha-stimulated NF-kappa B activation in Jurkat T cells and HL-60 cells was studied. TNF-alpha readily stimulated NF-kappa B activity in both cell lines as assayed by electrophoretic mobility shift assay and the use of a human immunodeficiency virus-chloramphenicol acetyltransferase reporter construct. However, TNF-alpha stimulation did not increase cell ceramide levels in either cell line. The exogenous addition of a short chain ceramide, N-acetylsphingosine, to Jurkat cells had no effect on NF-kappa B activity. When Jurkat T cells were exposed to the glucosylceramide synthase inhibitor, 1-phenyl-2-decanoylamino-3-morpholino-1-propanol, endogenous ceramide levels increased 4-fold. The increase in ceramide, however, did not result in NF-kappa B activation nor did it potentiate TNF-alpha or phorbol ester-stimulated activity. We conclude that TNF-alpha-induced NF-kappa B activation occurs in Jurkat and HL-60 cell lines that do not demonstrate an increase in TNF-alpha-induced ceramide. Increasing ceramide levels by the addition of short chain ceramides or the use of a glucosylceramide synthase inhibitor can be dissociated from activation of NF-kappa B by TNF-alpha.
Betts
J C
JC
Department of Internal Medicine, University of Michigan, Ann Arbor 48109-0676.
Agranoff
A B
AB
Nabel
G J
GJ
Shayman
J A
JA
D000970:Antineoplastic Agents / Q000494:pharmacology; D002460:Cell Line; D002518:Ceramides / Q000378:metabolism*; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000378:metabolism; D005822:Genetic Vectors; D006678:HIV; D006801:Humans; D007700:Kinetics; D015473:Leukemia, Promyelocytic, Acute; D015458:Leukemia, T-Cell; D009025:Morpholines / Q000494:pharmacology; D016328:NF-kappa B / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D000970:Antineoplastic Agents; D002518:Ceramides; D009025:Morpholines; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; C033110:RV 538; D015500:Chloramphenicol O-Acetyltransferase; D013755:Tetradecanoylphorbol Acetate