Abstract
Malignant gliomas are diffusively infiltrative and remain among the deadliest of all cancers. NF-κB is a transcription factor that mediates cell growth, migration and invasion, angiogenesis and resistance to apoptosis. Normally, the activity of NF-κB is tightly regulated by numerous mechanisms. However, in many cancers, NF-κB is constitutively activated and may function as a tumor promoter. Herein, we show that in gliomas, NF-κB is constitutively activated and the levels of cIAP2, Bcl-2, Bcl-xL and Survivin are elevated. These genes are regulated by NF-κB and can inhibit apoptosis. To understand the potential role of NF-κB p65 in suppressing apoptosis, we generated human glioma cell lines that inducibly express shRNA molecules specific for p65. We demonstrate that in the absence of p65, TNF-α induced cIAP2 expression is significantly reduced while the levels of Bcl-2, Bcl-xL and Survivin are not affected. These data suggest that of these genes, only cIAP2 is a direct target of p65, which was confirmed using RT-PCR and chromatin immunoprecipitation (ChIP) assays. By reducing the levels of p65 and/or cIAP2 levels, we demonstrate that the levels of RIP poly-ubiquitination are reduced, and that p65-deficient glioma cells are more sensitive to the cytotoxic effects of TNF-α. Specifically, in the presence of TNF-α glioma cells lacking p65 and/or cIAP2 showed cellular proliferation defects and underwent cell death. These data suggest that NF-κB and/or cIAP2 may be therapeutically relevant targets for the treatment of malignant gliomas.
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Acknowledgments
This work was supported in part by Public Service Grants CA-97247 from the National Cancer Institute (E. N. B.), NS-50665 from the National Institute of Neurological Disorders and Stroke (E. N. B.), CA-13148-35 from the National Cancer Institute (E. N. B.), 5P30CA013149-38 from the UAB Comprehensive Cancer Center (E. N. B.), IRG-60-001-47 from the American Cancer Society (S. N.), CA-13148-31 from the National Cancer Institute (S. N.), and funding from the Southeastern Brain Tumor Foundation (S. N.). We thank Dr. Xinbin Chen for providing the pBABE-HI-TetO plasmid, Dr. Tong Zhou for providing antibodies specific for Bcl-2 and Bcl-xL, and Dr. G. Yancey Gillespie and the UAB Brain Tumor Tissue Core for providing the brain tissue samples used herein. We also thank Maria G. Salazar of the CFAR/CCC DNA Sequencing Core at UAB for expert advice and technical assistance.
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Xueyan Zhao and Travis Laver contributed equally to this work.
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Zhao, X., Laver, T., Hong, S.W. et al. An NF-κB p65-cIAP2 link is necessary for mediating resistance to TNF-α induced cell death in gliomas. J Neurooncol 102, 367–381 (2011). https://doi.org/10.1007/s11060-010-0346-y
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DOI: https://doi.org/10.1007/s11060-010-0346-y