V843I, a Lung Cancer Predisposing EGFR Mutation, Is Responsible for Resistance to EGFR Tyrosine Kinase Inhibitors
V843I, a Lung Cancer Predisposing EGFR Mutation, Is Responsible for Resistance to EGFR Tyrosine Kinase Inhibitors
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DOI：
10.1097/jto.0000000000000241
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发表时间：
2014-09-01
影响因子：
20.4
通讯作者：
Watanabe, Takashi
中科院分区：
文献类型：
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作者：
Matsushima, Satsuki;Ohtsuka, Kouki;Watanabe, Takashi
简介：我们以前证明，一个家庭倾向于肺癌窝藏V843 I取代表皮生长因子受体（EGFR）蛋白。我们在这里报告的进一步表征突变EGFR蛋白的致瘤性和耐药的背景下，酪氨酸激酶抑制剂（TKIs）的EGFR activity.Methods：磷酸化的EGFR和下游信号蛋白的肺腺癌细胞系EGFR突变的流式细胞术进行了测定。使用细胞活力试验研究了这些细胞系对TKI的敏感性，无论是否通过小抑制性RNA（siRNA）抑制突变型EGFR表达。此外，蛋白质建模被用来预测TKI结合携带V843 I突变的EGFR蛋白。结果：EGFR和下游信号蛋白的磷酸化升高后，转染与V843 I的EGFR基因。尽管具有V843 I + L 858 R的细胞系表现出对EGFR-TKI的抗性，但在与V843 I等位基因特异性的siRNA孵育后，细胞变得对TKI敏感。结论：V843 I突变通过促进EGFR及其下游信号蛋白磷酸化参与肿瘤的发生。该突变似乎还通过EGFR的结构修饰提供对EGFR-TKI的耐药性。这些特征与EGFR T790 M突变的特征相似，表明具有种系V843 I或T790 M突变的病例可归类为一类对EGFR-TKI耐药的家族性肺癌综合征。
Introduction: We previously demonstrated that a family predisposed to lung cancer harbored a V843I substitution in the epidermal growth factor receptor (EGFR) protein. We report here the further characterization of this mutant EGFR protein in the context of tumorigenicity and resistance to tyrosine kinase inhibitors (TKIs) of EGFR activity.Methods: Phosphorylation of EGFR and downstream signaling proteins of lung adenocarcinoma cell lines with EGFR mutations was assayed by flow cytometry. Susceptibility to TKIs of these cell lines, with or without suppression of mutant EGFR expression by small inhibitory RNA (siRNA), was investigated using a cellular viability assay. Furthermore, protein modeling was used to predict TKI binding to EGFR protein carrying the V843I mutation.Results: Phosphorylation of EGFR and downstream signaling proteins was elevated upon transfection with an EGFR gene with the V843I. Although the cell line with V843I + L858R demonstrated resistance to EGFR-TKIs, the cells became susceptible to TKIs upon incubation with siRNA specific for the V843I allele. The structural analysis suggested that TKI binding to EGFR would be sterically hindered by Arg841 in the double-mutant (V843I + L858R) EGFR.Conclusions: The V843I mutation contributes to tumorigenesis by promoting phosphorylation of EGFR and its downstream signaling proteins. This mutation also appears to provide resistance to EGFR-TKIs through structural modification of EGFR. These features are comparable with those in EGFR T790M mutation, suggesting that cases with germ-line V843I or T790M mutations could be categorized as a class of familial lung cancer syndrome with resistance to EGFR-TKIs.