Most T790M mutations are present on the same EGFR allele as activating mutations in patients with non-small cell lung cancer
Most T790M mutations are present on the same EGFR allele as activating mutations in patients with non-small cell lung cancer
复制标题
DOI：
10.1016/j.lungcan.2017.02.019
复制
发表时间：
2017-06-01
期刊：
影响因子：
5.3
通讯作者：
Nakanishi, Yoichi
中科院分区：
文献类型：
--
作者：
Hidaka, Noriko;Iwama, Eiji;Nakanishi, Yoichi
目的：表皮生长因子受体基因（EGFR）的T790 M和C797 S突变分别使携带EGFR激活突变的非小细胞肺癌（NSCLC）患者对第一代和第三代EGFR酪氨酸激酶抑制剂（TKI）耐药。在第一代和第三代EGFR-TKI治疗失败的患者的肿瘤标本中，已发现C797 S与T790 M呈顺式或反式。然而，T790M和EGFR激活突变之间的等位基因关系尚未得到很好的表征。我们现在已经开发了一种基于数字聚合酶链反应（dPCR）的方法，用于确定两种EGFR突变之间的等位基因关系（T790M和C797S或活化突变）。用这种新方法分析了7个临床NSCLC标本和2个同时携带激活突变和T790 M的NSCLC细胞系，以确定这些EGFR之间的等位基因关系。结果：激活突变和T790 M均阳性的等位基因数与T790 M阳性等位基因数的中位数比为97.1%（范围，90.0 - 100%）。通过下一代测序进行的确证性分析产生了96.7%的相应值（范围，89.1 - 99.5%）。因此，我们的dPCR方法可靠地确定了两个EGFR突变之间的等位基因关系，在定量martens.Conclusions：几乎所有的T790 M突变检测顺式与EGFR的激活突变，无论从头或获得的状态T790 M，与癌细胞窝藏T790 M和激活突变的同一等位基因出现EGFR-TKI治疗过程中选择和富集。(C)2017 Elsevier B.V.版权所有。
Objectives: The T790M and C797S mutations of the epidermal growth factor receptor gene (EGFR) confer resistance to first- and third-generation EGFR tyrosine kinase inhibitors (TKIs), respectively, in patients with non-small cell lung cancer (NSCLC) harboring activating mutations of EGFR. C797S has been identified in cis or in trans with T790M in tumor specimens from patients who experienced treatment failure with first- and third-generation EGFR-TKIs. The allelic relation between T790M and activating mutations of EGFR has not been well characterized, however. We have now developed a digital polymerise chain reaction (dPCR)-based method for determination of the allelic relation between two types of EGFR mutation (T790M and either C797S or an activating mutation).Materials and Methods: Seven clinical NSCLC specimens and two NSCLC cell lines harboring both an activating mutation and T790M were analyzed with this new method to identify the allelic relation between these EGFR mutations.Results: The median ratio of the number of alleles positive for both an activating mutation and T790M to the number of T790M-positive alleles was 97.1% (range, 90.0-100%). Confirmatory analysis by next-generation sequencing yielded a corresponding value of 96.7% (range, 89.1-99.5%). Our dPCR method thus reliably identifies the allelic relation between two EGFR mutations in a quantitative manner.Conclusions: Almost all T790M mutations were detected in cis with activating mutations of EGFR regardless of the de novo or acquired status of T790M, with cancer cells harboring T790M and activating mutations on the same allele appearing to be selected and enriched during EGFR-TKI treatment. (C) 2017 Elsevier B.V. All rights reserved.