title: The prevention of the staurosporine-induced apoptosis by Bcl-X(L), but not by Bcl-2 or caspase inhibitors, allows the extensive differentiation of human neuroblastoma cells.
abstract: Staurosporine is one of the best apoptotic inducers in different cell types including neuroblastomas. In this study we have compared the efficiency and final outcome of three different anti-apoptotic strategies in staurosporine-treated SH-SY5Y human neuroblastoma cells. At staurosporine concentrations up to 500 nm, z-VAD.fmk a broad-spectrum, noncompetitive inhibitor of caspases, reduced apoptosis in SH-SY5Y cells. At higher concentrations, z-VAD.fmk continued to inhibit caspases and the apoptotic phenotype but not cell death which seems to result from oxidative damage. Stable over-expression of Bcl-2 in SH-SY5Y protected cells from death at doses of staurosporine up to 1 microm. At higher doses, cytochrome c release from mitochondria occurred, caspases were activated and cells died by apoptosis. Therefore, we conclude that Bcl-2 increased the threshold for apoptotic cell death commitment. Over-expression of Bcl-X(L) was far more effective than Bcl-2. Bcl-X(L) transfected cells showed a remarkable resistance staurosporine-induced cytochrome c release and associated apoptotic changes and survived for up to 15 days in 1 microm staurosporine. In these conditions, SH-SY5Y displayed a remarkable phenotype of neuronal differentiation as assessed by neurite outgrowth and expression of neurofilament, Tau and MAP-2 neuronal specific proteins.
authors.affiliation: Grup de Neurobiologia Molecular, Departament de Ciències Mèdiques Bàsiques, Universitat de Lleida, Lleida, Spain.
authors.forename: Víctor J
authors.initials: VJ
authors.lastname: Yuste
authors.forename: Isabel
authors.initials: I
authors.lastname: Sánchez-López
authors.forename: Carme
authors.initials: C
authors.lastname: Solé
authors.forename: Mario
authors.initials: M
authors.lastname: Encinas
authors.forename: Jose R
authors.initials: JR
authors.lastname: Bayascas
authors.forename: Jacint
authors.initials: J
authors.lastname: Boix
authors.forename: Joan X
authors.initials: JX
authors.lastname: Comella
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D061945:Caspase Inhibitors; D002454:Cell Differentiation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D016501:Neurites / Q000187:drug effects / Q000473:pathology / Q000502:physiology; D009447:Neuroblastoma / Q000473:pathology* / Q000503:physiopathology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000494:pharmacology*; D019311:Staurosporine / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D051020:bcl-X Protein
chemical_list: D000590:Amino Acid Chloromethyl Ketones; C494811:BCL2L1 protein, human; D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; D019253:Proto-Oncogene Proteins c-bcl-2; D051020:bcl-X Protein; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D019311:Staurosporine