Molecular Diagnosis of Activating EGFR Mutations in Non-Small Cell Lung Cancer Using Mutation-Specific Antibodies for Immunohistochemical Analysis
Molecular Diagnosis of Activating EGFR Mutations in Non-Small Cell Lung Cancer Using Mutation-Specific Antibodies for Immunohistochemical Analysis
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DOI：
10.1158/1078-0432.ccr-09-3239
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发表时间：
2010-06-15
影响因子：
11.5
通讯作者：
Ono, Mayumi
中科院分区：
文献类型：
--
作者：
Kawahara, Akihiko;Yamamoto, Chizuko;Ono, Mayumi
目的：非小细胞肺癌(NSCLC)对表皮生长因子受体(EGFR)靶向药物吉非替尼(Gefitinib)和厄洛替尼(Erlotinib)的疗效与激活EGFR突变密切相关。最常见的突变是外显子19的delE746-A750和外显子21的L858R，占所有EGFR突变的90%。最近，针对EGFR突变的特异性抗体被开发出来，并在免疫组织化学分析中表现良好，其敏感度接近90%。我们已经研究了这种方法检测激活的EGFR突变是否具有与直接DNA测序相当的灵敏度，直接DNA测序用于检测NSCLC中的这些突变。实验设计：我们使用针对外显子19的E746-A750缺失突变和外显子21的L858R点突变的抗体，在Western印迹分析和免疫组织化学中确定这些突变在NSCLC细胞系中的存在。结果：用抗delE746-A750抗体和抗L858R抗体在NSCLC肿瘤标本中检测EGFR突变的敏感性分别为79%和83%。结论：这种简便、快速的检测EGFR突变的方法，即使在IV期NSCLC患者的小支气管活检中也是如此，将有助于诊断NSCLC患者对EGFR靶向药物的反应。建议将其与DNA测序相结合，以开发改进的个性化EGFR靶向疗法。临床癌症资源；16(12)；3163-70。(C)2010年AACR。
Purpose: Therapeutic responses of non-small cell lung carcinoma (NSCLC) to epidermal growth factor receptor (EGFR)-targeted drugs, such as gefitinib and erlotinib, are closely associated with activating EGFR mutations. The most common mutations are delE746-A750 in exon 19 and L858R in exon 21, accounting for similar to 90% of all EGFR mutations. Recently, EGFR mutation-specific antibodies were developed and did well in immunohistochemical analysis, giving a sensitivity of similar to 90%. We have investigated whether this method detects activating EGFR mutations with sensitivity comparable with direct DNA sequencing, which is used to detect these mutations in NSCLC.Experimental Design: We used antibodies specific for the E746-A750 deletion mutation in exon 19 and the L858R point mutation in exon 21 in Western blot analysis and immunohistochemistry to determine the presence of these mutations in NSCLC cell lines. We also examined these EGFR mutations in NSCLC tumor samples from 60 patients by immunohistochemically and direct DNA sequencing.Results: We were able to identify EGFR mutations in NSCLC tumor samples immunohistochemically with a sensitivity of 79% using the anti-delE746-A750 antibody and 83% using the anti-L858R antibody. Additional DNA sequencing markedly improved the sensitivity obtained by immunohistochemistry.Conclusions: This simple and rapid assay for detecting EGFR mutations, even in the small bronchial biopsies obtained in stage IV NSCLC patients, will be useful for diagnosing responsiveness to EGFR-targeted drugs in patients with NSCLC. Combining this with DNA sequencing is recommended for the development of improved personalized EGFR-targeted therapeutics. Clin Cancer Res; 16(12); 3163-70. (C)2010 AACR.