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Intracellular phosphorylation of cis-4-methylsphingosine was previously shown to result in a metabolically stable compound that accumulates in Swiss 3T3 fibroblasts and mimics the mitogenic effect induced by the shortlived sphingosine metabolite, sphingosine 1-phosphate. In the present study incubation of neuroblastoma B104 cells with cis-4-methylsphingosine (10 M) also resulted in an intracellular accumulation of its phosphorylated derivative that was, however, associated with the concentrationdependent induction of apoptosis, not observed after treatment with 10 M of sphingosine-1-phosphate or sphingosine, respectively. In B104 cells, cis-4-methylsphingosine stimulated p38 mitogenactivated protein kinase (p38 MAPK) and simultaneously inhibited extracellular signalregulated kinase (ERK), whereas sphingosine and sphingosine 1-phosphate only stimulated p38 MAPK without suppression of ERK. Inhibition of cis-4-methylsphingosine phosphorylation reduced both, apoptosis and concurrent regulation of mitogenactivated protein kinases (MAPKs), suggesting that the unusual accumulation of the phosphorylated sphingoid base was responsible for the biological effects. Furthermore, inhibition of p38 MAPK prevented cis-4-methylsphingosineinduced apoptosis, while suppression of the ERK pathway in the presence of sphingosine or sphingosine-1-phosphate resulted in apoptosis, indicating that the simultaneous opposite regulation of the two MAPKs was required for the induction of apoptosis.

cis-4-Methylsphingosine Phosphate Induces Apoptosis in Neuroblastoma Cells by Opposite Effects on p38 and ERK Mitogen-Activated Protein Kinases