Shi Huiyang, Li Weihua, Guo Lei, Ying Jianming, Hu Xingsheng
Department of Medical Oncology, State Key Laboratory of Molecular Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
J Thorac Dis. 2025 Aug 31;17(8):5588-5596. doi: 10.21037/jtd-24-1787. Epub 2025 Aug 28.
Both amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) and next-generation sequencing (NGS) are able to identify sensitizing epidermal growth factor receptor () mutations of non-small cell lung cancer (NSCLC) patients. This study aims to determine the difference between ARMS-PCR and NGS among NSCLC patients with concurrent L858R and uncommon exon 21 mutations, as well as their treatment response to EGFR tyrosine kinase inhibitors (EGFR-TKIs).
Concurrent NGS and ARMS-PCR were performed with formalin-fixed, paraffin-embedded (FFPE) tumor tissues obtained from postoperative specimens, lung biopsy or fiberoptic bronchoscopy to identify mutations in 5,033 NSCLC cases. The clinical and pathological information of the study population was retrospectively collected from electronic clinical records at Cancer Hospital, Chinese Academy of Medical Sciences and followed up by phone calls or text messages. The mutant allele frequencies (MAFs) between L858R and uncommon exon 21 mutations were analyzed using Spearman correlation analysis and the survival information was analyzed using the Kaplan-Meier method and Log-rank tests.
Totally, L858R mutations were identified in 1,024 (20.3%) cases, among which 54 concurrent L858R and uncommon exon 21 mutations were identified by NGS but not ARMS-PCR. The most common concurrent pathogenic mutations in exon 21 were V834L, K860I and L833V. Of those, 23 patients received EGFR-TKIs as the first-line treatment with median progression-free survival (PFS) of 16.7 months [95% confidence interval (CI): 10.1-50.8 months], including 15 patients treated with first-generation EGFR-TKIs, 2 patients treated with second-generation EGFR-TKIs and 6 patients treated with third-generation EGFR-TKIs. Interestingly, concurrent L858R and K860I mutations were observed in 10 cases by NGS, while ARMS-PCR showed completely negative results in mutations. Of those, three patients received EGFR-TKIs as the first-line treatment, with median PFS of 11.0 months and ORR of 100.0%. Furthermore, three patients with concurrent L858R and K860I mutations identified by NGS received adjuvant targeted therapy after surgery with median disease-free survival of 10.0 months.
NSCLC patients with concurrent L858R and K860I mutations are able to benefit from EGFR-TKIs, which can be detected by NGS but not by ARMS-PCR. Thus, NGS should be recommended for NSCLC patients to identify driver alterations such as mutations, especially when negative results are obtained by ARMS-PCR.
扩增阻滞突变系统-聚合酶链反应（ARMS-PCR）和下一代测序（NGS）均能够识别非小细胞肺癌（NSCLC）患者的表皮生长因子受体（EGFR）敏感突变。本研究旨在确定在伴有L858R和罕见的21外显子突变的NSCLC患者中，ARMS-PCR和NGS之间的差异，以及他们对EGFR酪氨酸激酶抑制剂（EGFR-TKIs）的治疗反应。
对从术后标本、肺活检或纤维支气管镜检查获取的福尔马林固定、石蜡包埋（FFPE）肿瘤组织同时进行NGS和ARMS-PCR，以识别5033例NSCLC病例中的EGFR突变。研究人群的临床和病理信息从中国医学科学院肿瘤医院的电子临床记录中进行回顾性收集，并通过电话或短信进行随访。使用Spearman相关分析分析L858R和罕见的21外显子突变之间的突变等位基因频率（MAF），并使用Kaplan-Meier方法和Log-rank检验分析生存信息。
总共在1024例（20.3%）病例中鉴定出L858R突变，其中通过NGS鉴定出54例同时存在L858R和罕见的21外显子突变，但ARMS-PCR未检测到。21外显子中最常见的同时存在的致病性突变是V834L、K860I和L833V。其中，23例患者接受EGFR-TKIs作为一线治疗，中位无进展生存期（PFS）为16.7个月[95%置信区间（CI）：10.1 - 50.8个月]，包括15例接受第一代EGFR-TKIs治疗的患者、2例接受第二代EGFR-TKIs治疗的患者和6例接受第三代EGFR-TKIs治疗的患者。有趣的是，通过NGS在10例病例中观察到同时存在L858R和K860I突变，而ARMS-PCR在EGFR突变检测中显示完全阴性结果。其中，3例患者接受EGFR-TKIs作为一线治疗，中位PFS为11.0个月，客观缓解率（ORR）为100.0%。此外，3例通过NGS鉴定出同时存在L858R和K860I突变的患者术后接受辅助靶向治疗，中位无病生存期为10.0个月。
同时存在L858R和K860I突变的NSCLC患者能够从EGFR-TKIs中获益，这些突变可通过NGS检测到，但不能通过ARMS-PCR检测到。因此，对于NSCLC患者，应推荐使用NGS来识别驱动改变，如EGFR突变，尤其是当ARMS-PCR结果为阴性时。
