Background: Epidermal growth factor receptor (EGFR) gene mutation is closely related to the EGFR-TKI target treatment and prognosis of lung adenocarcinoma patients. The mutation status of EGFR is limited by tissue detection. The purpose of this study was to investigate the difference of EGFR mutants in plasmacirculating cell-free DNA (cfDNA) obtained from patients with non-small cell lung cancer (NSCLC) in three groups: pre-therapy, after traditional chemotherapy and targeted therapy. The aim of this study was to analyze whether the plasma cfDNA could effectively determine the EGFR mutations and monitor the drug resistant gene T790M, as well as its prognostic prediction value in patients with targeted therapy.
Methods: ARMS (amplification refractory mutation system)-PCR was used to detect EGFR mutations in 107 (50 of pre-therapy, 29 after traditional chemotherapy and 28 after targeted therapy) cases of paired plasma and tumor tissue specimens, followed by comparing their concordance. The sensitivity, specificity and the prognostic value of plasma cfDNA detection were also observed.
Results: The total rate of EGFR mutation was 56% (60/107) in all plasma samples and 77.6% (83/107) in corresponding tumor tissues. Completely the same mutants and wild-type EGFR were found in 68.2% cases of paired specimens. The sensitivity of plasma cfDNA detection was 72.3% and the specificity was up to 100%. Patients were sub-categorized according to therapy. The results showed that the highest consistent rate of cfDNA and tumor tissues was found in the group of pre-therapy (74%, 37/50). Whereas, the lowest consistent rate was observed in the targeted therapy group (57.1%, 16/28). It indicated that the targeted treatment could change the EGFR status in plasma cfDNA. Further analyses on inconsistent cases in this group revealed that 50% of them were compound EGFR mutations with T790M. Thereby, it suggested that targeted therapy might induce the emergence of drug resistance gene T790M. This speculation was confirmed by survival analyses. Based on plasma cfDNA results, patients with T790M mutant had significantly worse progression-free survival (PFS) and overall survival (OS).
Conclusions: For EGFR testing, ARMS-PCR on plasma cfDNA is a promising methodology with the highest specificity and effective sensitivity. It is useful for EGFR testing in patients before treatment, especially the late-stage patients. Simultaneously, plasma cfDNA could be used to monitor the drug resistant mutation, T790M status and predict prognosis after targeted therapy.
【中文题目：不同治疗对肺癌患者血液循环游离DNA 检测EGFR基因突变的影响及预后分析】 【中文摘要：背景与目的 表皮生长因子受体（epidermal growth factor receptor, EGFR）基因突变与肺腺癌患者TKI靶向治疗疗效和预后密切相关，常规组织分析其突变状态有诸多局限。本研究旨在探讨非小细胞肺癌（non-small cell lung cancer, NSCLC）患者血液循环游离DNA（cell-free DNA, cfDNA）检测EGFR基因突变在治疗前、传统化疗以及靶向治疗后的表达差异。分析血液cfDNA是否能准确检测EGFR基因突变并监测耐药基因T790M的变化，以及TKI在靶向治疗患者中的预后价值。方法 应用ARMS（amplification refractory mutation system）法检测107例（50例治疗前、29例传统化疗和28例靶向治疗）肺癌患者配对血液和肿瘤组织样本的EGFR基因突变并比较其表达差异；计算检测的一致性、敏感性和特异性；分析血检对靶向治疗患者的预后价值。结果 血浆cfDNA检测EGFR总突变率在107例肺癌患者中为56%（60例），而配对肿瘤组织样本检出率为77.6% （83例）。一一配对比较发现两者总体一致率为68.2%。血检的敏感性是72.3%，特异性为100%。依据治疗状态分组后发现治疗前组患者血液和肿瘤组织样本的检测一致率最高（74%, 37/50），而靶向组一致率最低（57.1%, 16/28），提示靶向治疗改变血浆cfDNA中EGFR基因状态。具体分析靶向组不一致病例发现50%新检出含T790M的双突变，提示靶向治疗后耐药基因出现。生存分析证实血检含T790M双突变组的无进展生存期（progression-free survival, PFS）和总生存期（overall survival, OS）均显著低于无T790M突变组。结论 应用ARMS法检测血液循环游离DNA（circulating cell-free DNA, cfDNA）的EGFR基因突变是一种特异性高、敏感性好的检测方法。适用于治疗前晚期肺癌患者的EGFR基因突变状态检测。同时，适用于靶向治疗后监测T790M耐药突变状态及预测患者预后。 】 【中文关键词：肺腺癌；循环游离DNA；EGFR；T790M突变；预后】.
Keywords: Circulating cell-free DNA; EGFR; Lung neoplasms; Prognosis; T790M.