Zhao Jing, Feng Hua-Hua, Zhao Jin-Yin, Liu Li-Cheng, Xie Fei-Fei, Xu Yan, Chen Min-Jiang, Zhong Wei, Li Long-Yun, Wang Han-Ping, Zhang L I, Xiao Y I, Chen Wei-Jun, Wang Meng-Zhao
Department of Respiratory Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730, P.R. China.
Beijing BGI-GBI Biotech Co., Ltd., Beijing 101300, P.R. China.
Oncol Lett. 2016 Apr;11(4):2573-2579. doi: 10.3892/ol.2016.4263. Epub 2016 Feb 23.
The current study aimed to develop a method to rapidly, sensitively and practically screen for the epidermal growth factor receptor (EGFR) T790M mutation. This method combines an allele-specific competitive blocker (ACB) with a TaqMan quantitative polymerase chain reaction (PCR) amplification refractory mutation system (ARMS) in a one-step reaction. Using a mimic of a human genomic DNA panel containing serially diluted mutant alleles, the performance efficacy of this method was assessed. Using this method, the EGFR T790M mutation was detected in tyrosine kinase inhibitor (TKI)-naïve samples obtained from 27 non-small cell lung cancer (NSCLC) patients with EGFR-activating mutations. The association between T790M mutations and the clinical benefit of EGFR-TKI treatment was also analysed. The sensitivity of this method was as low as 0.01%. In the samples from the 27 NSCLC patients, this method identified 6 mutant patients (22.2%), which was higher than the detection rate with scorpion ARMS (0.0%). No clinical variables were associated with the occurrence of a T790M mutation. The median progression-free survival time in the TKI-naïve patients with a T790M mutation was shorter that that of patients without the mutation, but the difference was not significant (3.2 vs. 19.5 months, respectively; P=0.256). The median overall survival time in the groups with or without T790M mutation also did not significantly differ (10 vs. 20 months, respectively; P=0.689). Overall, the ACB-ARMS PCR method could be useful for detecting the EGFR T790M mutation in clinical samples that contain only a small number of mutant alleles. The clinical significance of a T790M mutation should be further investigated.
本研究旨在开发一种快速、灵敏且实用的方法，用于筛查表皮生长因子受体（EGFR）T790M突变。该方法将等位基因特异性竞争阻断剂（ACB）与TaqMan定量聚合酶链反应（PCR）扩增阻滞突变系统（ARMS）结合在一步反应中。使用包含连续稀释突变等位基因的人类基因组DNA模板模拟物，评估了该方法的性能效果。使用该方法，在从27例具有EGFR激活突变的非小细胞肺癌（NSCLC）患者中获取的未经酪氨酸激酶抑制剂（TKI）治疗的样本中检测到了EGFR T790M突变。还分析了T790M突变与EGFR-TKI治疗临床获益之间的关联。该方法的灵敏度低至0.01%。在27例NSCLC患者的样本中，该方法鉴定出6例突变患者（22.2%），高于蝎形ARMS的检测率（0.0%）。没有临床变量与T790M突变的发生相关。T790M突变的未经TKI治疗患者的无进展生存期（PFS）中位数短于无该突变的患者，但差异无统计学意义（分别为3.2个月和19.5个月；P=0.256）。有或无T790M突变组的总生存期（OS）中位数也无显著差异（分别为10个月和20个月；P=0.689）。总体而言，ACB-ARMS PCR方法可用于检测仅含有少量突变等位基因的临床样本中的EGFR T790M突变。T790M突变的临床意义应进一步研究。
