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Originally published In Press as doi:10.1074/jbc.M203891200 on August 26, 2002

J. Biol. Chem., Vol. 277, Issue 44, 42144-42150, November 1, 2002
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Constitutively Active NFkappa B Is Required for the Survival of S-type Neuroblastoma*

Xin BianDagger §, Anthony W. Opipari Jr.§, Anthony B. RatanaproeksaDagger , Anthony E. Boitano||, Peter C. Lucas**, and Valerie P. CastleDagger Dagger Dagger

From the Departments of Dagger  Pediatrics,  Obstetrics and Gynecology, || Chemistry, and ** Pathology, University of Michigan, Ann Arbor, Michigan 48109

The NFkappa B transcription factors can both promote cell survival and induce apoptosis depending on cell type and context. Neuroblastoma (NB) cells display two predominant culture phenotypes identified as N- and S-types. Malignant S-type cells express neither high levels of MYCN nor Bcl-2, suggesting that other survival mechanisms are important. We characterized NFkappa B activity in S-type cells and determined its role in their survival. S-type lines (SH-EP1 and SK-N-AS) were treated with pyrrolidine dithiocarbamate (PDTC), a NFkappa B inhibitor, or L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK), a serine protease inhibitor that blocks Ikappa Balpha degradation. Both agents induced cell death, suggesting that constitutive NFkappa B activity is required for survival. The transient expression of a super-repressor Ikappa Balpha mutant killed S-type cells. The inhibition of NFkappa B produced an apoptotic response characterized by the collapse of the mitochondrial transmembrane electrochemical gradient, caspase-9 activation, and apoptotic DNA changes. Constitutive NFkappa B DNA binding activity specifically involving p65 and p50 was demonstrated in S- but not N-type cells by electromobility supershift and gene reporter assays. This study demonstrates a role for NFkappa B in the survival of S-type NB tumor cells and suggests that NFkappa B activity and function differ according to NB tumor cell phenotype.

* This work was supported in part by Grant CA697276-04 from the National Institutes of Health, the Janette Ferrantino Hematology Research Fund (to V. P. C.), and the Ravitz Foundation (to A. W. O and V. P. C.).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.

§ Both authors contributed equally to this work.

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

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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