Radiation-induced TNFα cross signaling-dependent nuclear import of NFκB favors metastasis in neuroblastoma
Radiation-induced TNFα cross signaling-dependent nuclear import of NFκB favors metastasis in neuroblastoma
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DOI：
10.1007/s10585-013-9580-y
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发表时间：
2013-08-01
影响因子：
4
通讯作者：
Aravindan, Natarajan
中科院分区：
文献类型：
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作者：
Aravindan, Sheeja;Natarajan, Mohan;Aravindan, Natarajan
确定NF-κ B B对放射反应的功能特异性协调可能揭示神经母细胞瘤诱导复发和转移的分子蓝图。我们最近证实，电离辐射（IR）引发的TNF α-NF κ B反馈信号引起的NF κ B持续活化导致神经母细胞瘤的放射抗性和复发。我们研究了抑制IR触发的或TNF α依赖的第二信号反馈依赖的NF κ B核输入是否会限制IR改变的侵袭和转移。将神经母细胞瘤细胞暴露于2戈伊并孵育1 h或24 h。然后用NF κ B靶向肽阻断剂SN 50处理细胞。在确认DNA结合活性、转录驱动的NF kappa B反式激活和可溶性TNF α分泌的阻断后，通过QPCR分析评估了93个肿瘤侵袭/转移基因的转录改变，然后在蛋白质水平上选择性验证。2戈伊照射在存活的SH-SY 5 Y、IMR-32和SK-N-MC细胞中分别诱导了63、42和71个基因。阻断NF κ B的翻译后核输入全面抑制IR触发的基因的初始活化（62/63、34/42和65/71）以及TNF α介导的肿瘤侵袭和转移信号分子的第二信号依赖性持续活化（59/63、32/42和71/71）。此外，MMP 9、MMP 2、PYK-2、SPA-1、Dnmt 3 B、Ask-1、CTGF、MMP 10、MTA-2、NF-2、E-Cadherin、TIMP-2和ADAMTS 1蛋白的改变以及我们的划痕试验结果证实了翻译后NF κ B B在IR调节的侵袭/转移中的作用。这些数据表明，IR诱导的第二阶段（翻译后）NF κ B活化介导TNF α依赖性第二信号传导，并进一步暗示IR诱导的NF κ B在治疗后存活的细胞中调节肿瘤侵袭/转移信号传导。
Ascertaining function-specific orchestration of NF kappa B 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 NF kappa B caused by ionizing radiation (IR)-initiated TNF alpha-NF kappa B feedback signaling leads to radioresistance and recurrence of neuroblastoma. We investigated whether muting IR-triggered or TNF alpha-dependent second-signaling feedback-dependent NF kappa B 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 NF kappa B-targeting peptide blocker, SN50. Upon confirming the blockade in DNA-binding activity, transcription driven transactivation of NF kappa B and secretion of soluble TNF alpha, 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 NF kappa B comprehensively inhibited both initial activation of genes (62/63, 34/42 and 65/71) triggered by IR and also TNF alpha-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 NF kappa B in IR-regulated invasion/metastasis. These data demonstrate that IR-induced second-phase (post-translational) NF kappa B activation mediates TNF alpha-dependent second signaling and further implies that IR induced NF kappa B in cells that survive after treatment regulates tumor invasion/metastasis signaling.