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Papers In Press, published online ahead of print October 25, 2001
Department of Pediatrics Hematology/Oncology, University of Michigan, Ann Arbor, MI 48109-0938
Corresponding Author: vcastle{at}umich.edu
Neuroblastoma is the most common extracranial solid tumor of childhood. N-type neuroblastoma cells (represented by SH-SY5Y and IMR32 cell lines) are characterized by a neuronal phenotype. N-type cell lines are generally N-myc amplified, express the anti-apoptotic protein Bcl-2, and do not express caspase-8. The present study was designed to determine the mechanism by which N-type cells die in response to specific cytotoxic agents (cisplatin, doxorubicin) commonly used to treat this disease. We found that N-type cells were equally sensitive to cisplatin and doxorubicin. Yet death induced by cisplatin was inhibited by the non-selective caspase inhibitor z-Val-Ala-Asp-fluoromethylketone or the specific caspase-9 inhibitor Ac-Leu-Glu-His-Asp-CHO; whereas in contrast, caspase inhibition did not prevent doxorubicin-induced death. Neither reactive oxygen species nor the mitochondrial permeability transition appears to play important roles in this process. Doxorubicin induced NF-
J. Biol. Chem, 10.1074/jbc.M108674200
Submitted on September 7, 2001
Revised on October 10, 2001
Accepted on October 25, 2001
NF-
B activation mediates doxorubicin-induced cell death in N-type neuroblastoma cells
B transcriptional activation in association with I-B
degradation prior to loss of cell viability. Surprisingly, the antioxidant and NF-B inhibitor, pyrrolidine dithiocarbamate blocked doxorubicin-induced NF-B transcriptional activation and provided profound protection against doxorubicin killing. Moreover, SH-SY5Y cells expressing a super-repressor form of I-B were completely resistant to doxorubicin killing. Together these findings show that NF-B activation mediates doxorubicin-induced cell death without evidence of caspase function and suggest that cisplatin and doxorubicin engage different death pathways to kill neuroblastoma cells.
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