Abstract
The mechanism of the inhibitory effect of anti-GD2 ganglioside (GD2) 14G2a mouse monoclonal antibody (mAb) on human neuroblastoma cells survival was studied in vitro. It was recently shown in IMR-32 cells that death induced by this antibody exhibited several characteristics typical of apoptosis. In this study we used cytotoxixity assays, qRT-PCR and immunoblotting to evaluate the response of several human neuroblastoma cell lines to the anti-GD2 14G2a mAb. We showed that the mAb decreases all three aurora kinases expression and phosphorylation in IMR-32 and LA-N-1 cells. Most importantly, we show, that MK-5108 specific aurora A kinase inhibitor decreases neuroblastoma cell survival, and when used in combination with the mAb, significantly potentiates cytotoxicity against IMR-32, CHP-134, and LA-N-5 neuroblastoma cells in vitro. It was shown that downregulation of aurora A kinase by the therapeutic antibody is associated with decreased levels of MYCN protein in cytoplasm, and induced expression of PHLDA1 and P53 proteins.
Keywords:
Aurora kinases; GD2 ganglioside; Monoclonal antibody; Neuroblastoma; P53.
Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal / immunology
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Antibodies, Monoclonal / pharmacology*
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Aurora Kinase A / antagonists & inhibitors*
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Aurora Kinase A / genetics
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Aurora Kinase A / metabolism
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Caspase 3 / metabolism
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Caspase 7 / metabolism
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Cell Line, Tumor
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Cell Survival / drug effects
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Cell Survival / immunology
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Cyclohexanecarboxylic Acids / pharmacology*
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Flow Cytometry
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Gangliosides / antagonists & inhibitors*
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Gangliosides / immunology
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Gangliosides / metabolism
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Gene Expression / drug effects
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Humans
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Immunoblotting
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Mice
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N-Myc Proto-Oncogene Protein
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Neuroblastoma / genetics
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Neuroblastoma / metabolism
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Neuroblastoma / pathology
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Nuclear Proteins / metabolism
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Oncogene Proteins / metabolism
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Phosphorylation / drug effects
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Protein Kinase Inhibitors / pharmacology
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Reverse Transcriptase Polymerase Chain Reaction
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Thiazoles / pharmacology*
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Tumor Suppressor Protein p53 / genetics
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Tumor Suppressor Protein p53 / metabolism
Substances
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4-(3-chloro-2-fluorophenoxy)-1-((6-(1,3-thiazol-2-ylamino)pyridin to 2-yl)methyl) cyclohexanecarboxylic acid
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Antibodies, Monoclonal
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Cyclohexanecarboxylic Acids
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Gangliosides
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MYCN protein, human
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N-Myc Proto-Oncogene Protein
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Nuclear Proteins
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Oncogene Proteins
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PHLDA1 protein, human
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Protein Kinase Inhibitors
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TP53 protein, human
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Thiazoles
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Transcription Factors
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Tumor Suppressor Protein p53
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ganglioside, GD2
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Aurora Kinase A
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Caspase 3
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Caspase 7