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Originally published In Press as doi:10.1074/jbc.M108674200 on October 25, 2001

J. Biol. Chem., Vol. 276, Issue 52, 48921-48929, December 28, 2001
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NF-kappa B Activation Mediates Doxorubicin-induced Cell Death in N-type Neuroblastoma Cells*

Xin BianDagger , Linda M. McAllister-LucasDagger , Feng Shao§, Kurt R. SchumacherDagger , Zhiwei Feng, Alan G. Porter||, Valerie P. CastleDagger **, and Anthony W. Opipari Jr.Dagger Dagger

From the Departments of Dagger  Pediatrics, § Biological Chemistry, and Dagger Dagger  Obstetrics and Gynecology, University of Michigan Medical School, Ann Arbor, Michigan 48109-0938 and the  Institute of Molecular and Cell Biology, 30 Medical Drive, Singapore 117609, Republic of Singapore

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 (such as cisplatin and 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 nonselective caspase inhibitor z-Val-Ala-Asp-fluoromethylketone or the specific caspase-9 inhibitor N-acetyl-Leu-Glu-His-Asp-aldehyde, whereas in contrast, caspase inhibition did not prevent doxorubicin-induced death. Neither the reactive oxygen species nor the mitochondrial permeability transition appears to play an important role in this process. Doxorubicin induced NF-kappa B transcriptional activation in association with I-kappa Balpha degradation prior to loss of cell viability. Surprisingly, the antioxidant and NF-kappa B inhibitor pyrrolidine dithiocarbamate blocked doxorubicin-induced NF-kappa B transcriptional activation and provided profound protection against doxorubicin killing. Moreover, SH-SY5Y cells expressing a super-repressor form of I-kappa B were completely resistant to doxorubicin killing. Together these findings show that NF-kappa 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.

* This work was supported by Grant CA69276-04 from the National Institutes of Health, by funds from the Jeanette Ferrantino Hematology Research Fund (to V. P. C.), and by funds from the Institute of Molecular and Cell Biology (Singapore).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

|| An adjunct staff member of the Department of Surgery, National University of Singapore.

** To whom correspondence should be addressed: Rm. 4302, University of Michigan Comprehensive Cancer Center, 1500 East Medical Center Dr., Ann Arbor, MI 48109-0938. Tel.: 734-763-0019; Fax: 734-647-9654; E-mail: vcastle@umich.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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