title: Activation of caspase-3 alone is insufficient for apoptotic morphological changes in human neuroblastoma cells.
abstract: Activated caspase-3 is considered an important enzyme in the cell death pathway. To study the specific role of caspase-3 activation in neuronal cells, we generated a stable tetracycline-regulated SK-N-MC neuroblastoma cell line, which expressed a highly efficient self-activating chimeric caspase-3, consisting of the caspase-1 prodomain fused to the caspase-3 catalytic domain. Under expression-inducing conditions, we observed a time-dependent increase of processed caspase-3 by immunostaining for the active form of the enzyme, intracellular caspase-3 enzyme activity, as well as poly(ADP-ribose) polymerase (PARP) cleavage. Induced expression of the caspase fusion protein showed predominantly caspase-3 activity without any apoptotic morphological changes. In contrast, staurosporine treatment of the same cells resulted in activation of multiple caspases and profound apoptotic morphology. Our work provides evidence that auto-activation of caspase-3 can be efficiently achieved with a longer prodomain and that neuronal cell apoptosis may require another caspase or activation of multiple caspase enzymes.
authors.affiliation: Department of Cardiovascular Research, Eli Lilly and Company, Indianapolis, USA.
authors.forename: Margaret M
authors.initials: MM
authors.lastname: Racke
authors.forename: Marian
authors.initials: M
authors.lastname: Mosior
authors.forename: Steve
authors.initials: S
authors.lastname: Kovacevic
authors.forename: Chan Hsin S
authors.initials: CH
authors.lastname: Chang
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Glasebrook
authors.forename: Neal W
authors.initials: NW
authors.lastname: Roehm
authors.forename: Songqing
authors.initials: S
authors.lastname: Na
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000235:genetics / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D004318:Doxycycline / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology; D017434:Protein Structure, Tertiary / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D019311:Staurosporine / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D000900:Anti-Bacterial Agents; D004791:Enzyme Inhibitors; D011993:Recombinant Fusion Proteins; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D019311:Staurosporine; D004318:Doxycycline