NFκB Signaling Related Molecular Alterations in Human Neuroblastoma Cells after Fractionated Irradiation
NFκB Signaling Related Molecular Alterations in Human Neuroblastoma Cells after Fractionated Irradiation
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DOI：
10.1269/jrr.08110
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发表时间：
2009-07-01
影响因子：
2
通讯作者：
Aravindan, Natarajan
中科院分区：
文献类型：
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作者：
Madhusoodhanan, Rakhesh;Natarajan, Mohan;Aravindan, Natarajan
放射适应性反应/NF κ B/分次照射/神经母细胞瘤/FIR后的转录反应。放射治疗已被用作高危神经母细胞瘤的一种连续性局部控制方式。然而，由于放射抵抗而导致的复发影响了放射治疗的成功。在靶向分子治疗中，确定分次辐射（FIR）调制的分子靶点是必要的。因此，我们研究了（i）代表六种功能途径的基因表达;（ii）NF κ B B DNA结合活性和（iii）在人神经母细胞瘤细胞中单剂量（10戈伊）辐射（SDR）和FIR（2戈伊x 5）后放射应答分子的表达。使用QPCR分析基因表达的变化，使用电泳迁移率变动分析（EMSA）分析NF κ B活性，使用免疫印迹分析PI κ B α。使用定量实时PCR（Q-PCR）和免疫印迹法测定TNF α、IL-1 α pAKT、IAP 1、IAP 2、XIAP、存活素、MnSOD、BID、巴克、MyD 88和Vegfc的调节。与SDR相比，FIR显著诱导了25个基因的表达，并完全抑制了另外30个基因的表达。此外，FIR诱导NF κ B-DNA结合活性和I κ B α磷酸化。同样，我们观察到IAP 1、IAP 2、XIAP、Survivin的诱导表达。IL-1 α、MnSOD、Bid、巴克、MyD 88、TNF α和pAKT。研究结果清楚地表明SDR和FIR之间细胞分子反应的明显差异。我们确定了几个可能影响FIR后辐射抗性的NF κ B B信号级联的潜在靶点。
Radioadaptive response/NF kappa B/Fractionated irradiation/Neuroblastoma/Transcriptional responses after FIR. Radiotherapy has been used as an adjunctive local-control modality for high-risk neuroblastoma. However, relapse due to radioresistance affects the success of radiotherapy. Ascertaining the fractionated radiation (FIR) modulated molecular targets is imperative in targeted molecular therapy. Accordingly, we investigated the (i) expression of genes representing six functional pathways; (ii) NF kappa B DNA-binding activity and (iii) expression of radioresponsive molecules after single dose (10 Gy) radiation (SDR) and FIR (2 Gy x 5) in human neuroblastoma cells. Alterations in gene expression were analyzed using QPCR-profiling, NF kappa B activity using electrophoretic mobility shift assay (EMSA) and PI kappa B alpha using immunoblotting. Modulations in TNF alpha, IL-1 alpha pAKT, IAP1, IAP2, XIAP, survivin, MnSOD, BID, Bak, MyD88 and Vegfc were determined using quantitative real-time PCR (Q-PCR) and immunoblotting. Compared to SDR, FIR significantly induced the expression of 25 genes and completely suppressed another 30 genes. Furthermore, FIR induced NF kappa B-DNA-binding activity and I kappa B alpha phosphorylation. Similarly, we observed an induced expression of IAP1, IAP2, XIAP, Survivin. IL-I alpha, MnSOD, Bid, Bak, MyD88, TNFa and pAKT in cells exposed to FIR. The results of the study clearly show distinct differences in the molecular response of cells between SDR and FIR. We identifed several potential targets confining to NF kappa B signaling cascade that may affect radio-resistance after FIR.