Abstract
Ascertaining function-specific orchestration of NFκB in response to radiation may reveal a molecular blue-print that dictates induced relapse and metastasis of the neuroblastoma. We recently demonstrated that sustained activation of NFκB caused by ionizing radiation (IR)-initiated TNFα–NFκB feedback signaling leads to radioresistance and recurrence of neuroblastoma. We investigated whether muting IR-triggered or TNFα-dependent second-signaling feedback–dependent NFκB nuclear import results in limiting IR-altered invasion and metastasis. Neuroblastoma cells were exposed to 2 Gy and incubated for 1 h or 24 h. The cells were then treated with an NFκB-targeting peptide blocker, SN50. Upon confirming the blockade in DNA-binding activity, transcription driven transactivation of NFκB and secretion of soluble TNFα, transcriptional alterations of 93 tumor invasion/metastasis genes were assessed by using QPCR profiling and then were selectively validated at the protein level. Exposure to 2 Gy induced 63, 42 and 71 genes in surviving SH-SY5Y, IMR-32 and SK-N-MC cells, respectively. Blocking post-translational nuclear import of NFκB comprehensively inhibited both initial activation of genes (62/63, 34/42 and 65/71) triggered by IR and also TNFα-mediated second signaling–dependent sustained (59/63, 32/42 and 71/71) activation of tumor invasion and metastasis signaling molecules. Furthermore, alterations in the proteins MMP9, MMP2, PYK-2, SPA-1, Dnmt3b, Ask-1, CTGF, MMP10, MTA-2, NF-2, E-Cadherin, TIMP-2 and ADAMTS1 and the results of our scratch-wound assay validate the role of post-translational NFκB in IR-regulated invasion/metastasis. These data demonstrate that IR-induced second-phase (post-translational) NFκB activation mediates TNFα-dependent second signaling and further implies that IR induced NFκB in cells that survive after treatment regulates tumor invasion/metastasis signaling.
Similar content being viewed by others
References
Burgos-Tiburcio A et al (2011) Development of targeted therapy for squamous cell carcinomas of the head and neck. Expert Rev Anticancer Ther 11(3):373–386
He H et al (2011) The novel protein TSR2 inhibits the transcriptional activity of nuclear factor-kappaB and induces apoptosis. Mol Biol (Mosk) 45(3):496–502
Valera FC et al (2010) NF-kappaB expression predicts clinical outcome for nasal polyposis. Rhinology 48(4):408–441
Macha MA et al (2011) Guggulsterone (GS) inhibits smokeless tobacco and nicotine-induced NF-kappaB and STAT3 pathways in head and neck cancer cells. Carcinogenesis 32(3):368–380
McGinn CJ, Kinsella TJ (1992) The experimental and clinical rationale for the use of S-phase-specific radiosensitizers to overcome tumor cell repopulation. Semin Oncol 19(4 Suppl 11):21–28
Parisi MT et al (1999) Complications of cancer therapy in children: a radiologist’s guide. Radiographics 19(2):283–297
Aggarwal BB (2004) Nuclear factor-kappaB: the enemy within. Cancer Cell 6(3):203–208
Abal M et al (2006) Molecular pathology of endometrial carcinoma: transcriptional signature in endometrioid tumors. Histol Histopathol 21(2):197–204
Baeuerle PA, Baltimore D (1991) Hormonal control regulation of gene transcription. In: Cohen P, Foulkes JG (eds) Molecular aspects of cellular regulation. Elsevier/North Holland Biomedical Press, Amsterdam, pp 409–432
Lenardo MJ, Baltimore D (1989) NF-kappa B: a pleiotropic mediator of inducible and tissue-specific gene control. Cell 58(2):227–229
Orlowski RZ, Baldwin AS Jr (2002) NF-kappaB as a therapeutic target in cancer. Trends Mol Med 8(8):385–389
Prasad AV et al (1994) Activation of nuclear factor kappa B in human lymphoblastoid cells by low-dose ionizing radiation. Radiat Res 138(3):367–372
Mohan N, Meltz ML (1994) Induction of nuclear factor kappa B after low-dose ionizing radiation involves a reactive oxygen intermediate signaling pathway. Radiat Res 140(1):97–104
Aravindan N et al (2008) Curcumin inhibits NFkappaB mediated radioprotection and modulate apoptosis related genes in human neuroblastoma cells. Cancer Biol Ther 7(4):569–576
Aravindan N et al (2008) Alteration of apoptotic signaling molecules as a function of time after radiation in human neuroblastoma cells. Mol Cell Biochem 310(1–2):167–179
Piva R, Belardo G, Santoro MG (2006) NF-kappaB: a stress-regulated switch for cell survival. Antioxid Redox Signal 8(3–4):478–486
Veeraraghavan J et al (2011) Radiation-triggered tumor necrosis factor (TNF) alpha-NFkappaB cross-signaling favors survival advantage in human neuroblastoma cells. J biol chem 286(24):21588–21600
Lin YZ et al (1995) Inhibition of nuclear translocation of transcription factor NF-kappa B by a synthetic peptide containing a cell membrane-permeable motif and nuclear localization sequence. J Biol Chem 270(24):14255–14258
Aravindan N et al (2011) Irreversible EGFR Inhibitor EKB-569 targets low-LET gamma-radiation-triggered rel orchestration and potentiates cell death in squamous cell carcinoma. PLoS ONE 6(12):e29705
Veeraraghavan J et al (2011) Low-dose gamma-radiation-induced oxidative stress response in mouse brain and gut: regulation by NFkappaB-MnSOD cross-signaling. Mutat Res 718(1–2):44–55
Giagkousiklidis S et al (2005) Sensitization for gamma-irradiation-induced apoptosis by second mitochondria-derived activator of caspase. Cancer Res 65(22):10502–10513
Madhusoodhanan R et al (2009) NFkappaB Signaling Related Molecular Alterations in Human Neuroblastoma Cells after Fractionated Irradiation. J Radiat Res(Tokyo) 50(4):311–324
Armstrong MB et al (2006) Signaling from p53 to NF-kappaB determines the chemotherapy responsiveness of neuroblastoma. Neoplasia 8(11):967–977
Bian X et al (2001) NF-kappa B activation mediates doxorubicin-induced cell death in N-type neuroblastoma cells. J Biol Chem 276(52):48921–48929
Bian X et al (2002) Constitutively active NFkappa B is required for the survival of S-type neuroblastoma. J Biol Chem 277(44):42144–42150
van de Stolpe A et al (1994) 12-O-tetradecanoylphorbol-13-acetate- and tumor necrosis factor alpha-mediated induction of intercellular adhesion molecule-1 is inhibited by dexamethasone. Functional analysis of the human intercellular adhesion molecular-1 promoter. J biol chem 269(8):6185–6192
Thomsen LL, Miles DW (1998) Role of nitric oxide in tumour progression: lessons from human tumours. Cancer metastasis rev 17(1):107–118
Helbig G et al (2003) NF-kappaB promotes breast cancer cell migration and metastasis by inducing the expression of the chemokine receptor CXCR4. J biol chem 278(24):21631–21638
Fujioka S et al (2003) Function of nuclear factor kappaB in pancreatic cancer metastasis. Clin Cancer Res 9(1):346–354
Corey JM et al (2010) Patterning N-type and S-type neuroblastoma cells with Pluronic F108 and ECM proteins. J Biomed Mater Res Part A 93(2):673–686
Okamoto T et al (2002) S-nitrosothiols inhibit cytokine-mediated induction of matrix metalloproteinase-9 in airway epithelial cells. Am J Respir Cell Mol Biol 27(4):463–473
Chayka O et al (2009) Clusterin, a haploinsufficient tumor suppressor gene in neuroblastomas. J Natl Cancer Inst 101(9):663–677
Granchi D et al (2004) In vitro blockade of receptor activator of nuclear factor-kappaB ligand prevents osteoclastogenesis induced by neuroblastoma cells. Int J Cancer 111(6):829–838
Acknowledgments
The authors were supported by National Institutes of Health (COBRE-1P20GM103639-01), American Cancer Society (Grant ACS-IRG-05-066-01), research development funds from the Department of Radiation Oncology at University of Oklahoma Health Sciences Center to N. Aravindan and the National Aeronautics and Space Administration (NASA) Ground- Based Studies in Space Radiobiology, Grant NNX12- AC32G to M. Natarajan.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Aravindan , S., Natarajan, M., Herman, T.S. et al. Radiation-induced TNFα cross signaling-dependent nuclear import of NFκB favors metastasis in neuroblastoma. Clin Exp Metastasis 30, 807–817 (2013). https://doi.org/10.1007/s10585-013-9580-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10585-013-9580-y