In tumor cells regulation of DNA double strand break repair through EGF receptor involves both NHEJ and HR and is independent of p53 and K-Ras status
In tumor cells regulation of DNA double strand break repair through EGF receptor involves both NHEJ and HR and is independent of p53 and K-Ras status
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DOI：
10.1016/j.radonc.2011.05.046
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发表时间：
2011-10-01
影响因子：
5.7
通讯作者：
Kriegs, Matte
中科院分区：
文献类型：
--
作者：
Myllynen, Laura;Rieckmann, Thorsten;Kriegs, Matte
目的：探讨表皮生长因子受体(EGFR)是否可作为调节肿瘤细胞DNA双链断裂(DSB)修复的通用靶点。材料和方法：以人肿瘤细胞株A549、H1299和HeLa为实验对象，灵长类细胞株CV1、EGF、ARC和TGFα为靶点，分别用西妥昔单抗或厄洛替尼抑制EGFR的激活。用Gamma-H_2AX/53BP1联合免疫染色和非同源末端连接(NHEJ)和同源重组(HR)方法检测DSB修复：用流式细胞仪检测细胞周期分布，用Western blotting检测蛋白表达。结果：无论使用何种配体，EGFR激活都能刺激DSB修复和NHEJ。当EGFR信号被阻断时，这种刺激被取消。在所有被测试的细胞系中都发现了这一规律，无论它们的P53或K-RAS状态如何。结论：EGFR对DSB修复的调节涉及NHEJ和HR两条途径，且与p53和K-ras基因突变状态无关，多数肿瘤细胞株中也存在EGFR对DSB修复的调控。(C)2011爱思唯尔爱尔兰有限公司。保留所有权利。放射治疗与肿瘤学101(2011)147-151
Purpose: The purpose of this study was to examine whether the epidermal growth factor receptor (EGFR) may be used as a general target to modulate DNA double strand break (DSB) repair in tumor cells.Material and methods: Experiments were performed with human tumor cell lines A549, H1299 and HeLa and primate cell line CV1, EGF, ARC and TGF alpha were used for EGFR activation, cetuximab or erlotinib for inhibition. Overall DSB repair was assessed by gamma H2AX/53BP1 co-immunostaining and non-homologous end-joining (NHEJ) and homologous recombination (HR) by using NHEJ and HR reporter cells: cell cycle distribution was determined by flow cytometry and protein expression by Western blot.Results: EGFR activation was found to stimulate overall DSB repair as well as NHEJ regardless of the ligand used. This stimulation was abolished when EGFR signaling was blocked. This regulation was found for all cell lines tested, irrespective of their p53 or K-Ras status. Stimulation and inhibition of EGFR were also found to affect HR.Conclusions: Regulation of DSB repair by EGFR involves both the NHEJ and HR pathway, and appears to occur in most tumor cell lines regardless of p53 and K-Ras mutation status. (C) 2011 Elsevier Ireland Ltd. All rights reserved. Radiotherapy and Oncology 101 (2011) 147-151