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Objective To investigate the potential mechanisms of eell death after the treatment with ceramide.Methods MTT assay,DNA ladder,reporter assay,FACS and Western blot assay were employed to investigate the potential mechanisms of cell death after the treatment with C2-ceramide.Results A short-time treatment with C2-ceramide induced cell death,which was associated with p38 MAP kinase activation,but had no links with typical caspase activation or PARP degradation.Rather than caspase inhibitor,Inhibitor of p38 MAP kinase blocked cell death induced by a short-time treatment with ceramide(<12 h). However,inhibition of p38 MAP kinase could not block cell death induced by a prolonged treatment with ceramide(>12h). Moreover,incubation of cells with ceramide for a long time(>12h)increased subG1,but reduced S phase accompanicd by caspase-dependent and caspase-independent changes including NFκB activation.Conclusion Ceramide-induced cell apoptosis involves both caspase-dependent and-independent signaling pathway.Caspase-independent cell death occurring in a relatively early stage,which is mediated via p38 MAP kinase.can progress into a stage involving both caspase-dependent and -independent mechanisms accompanied by cell signaling of MAPKs and NFκB.
Objective To investigate the potential mechanisms of eell death after the treatment with ceramide. Methods MTT assay, DNA ladder, reporter assay, FACS and Western blot assay were employed to investigate the potential mechanisms of cell death after the treatment with C2-ceramide. Results A short-time treatment with C2-ceramide induced cell death, which was associated with p38 MAP kinase activation, but had no links with typical caspase activation or PARP degradation. Rather than caspase inhibitor, Inhibitor of p38 MAP kinase blocked cell death induced by a short However, inhibition of p38 MAP kinase could not block cell death induced by a prolonged treatment with ceramide (> 12h). Furthermore, incubation of cells with ceramide for a long time (> 12h) increased subG1, but reduced S phase accompanicd by caspase-dependent and caspase-independent changes including NFκB activation. Conclusion Ceramide-induced cell apoptosis involves both caspase-dependent and-independent sigma naling pathway. Caspase-independent cell death occurring in a relatively early stage, which is mediated via p38 MAP kinase. progression into a stage involving both caspase-dependent and -independent mechanisms accompanied by cell signaling of MAPKs and NFKB.