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Activation of caspase‐3 alone is insufficient for apoptotic morphological changes in human neuroblastoma cells
Author(s) -
Racke Margaret M.,
Mosior Marian,
Kovacevic Steve,
Chang Chan Hsin S.,
Glasebrook Andrew L.,
Roehm Neal W.,
Na Songqing
Publication year - 2002
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.0022-3042.2002.00787.x
Subject(s) - caspase , caspase 3 , poly adp ribose polymerase , microbiology and biotechnology , apoptosis , staurosporine , caspase 10 , caspase 8 , enzyme activator , biology , caspase 9 , caspase 2 , cell culture , chemistry , programmed cell death , enzyme , biochemistry , signal transduction , polymerase , kinase , protein kinase a , genetics
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.