晚期非小细胞肺癌患者表皮生长因子受体基因突变检测

Detection of epidermal growth factor receptor gene mutations in advanced non-small cell lung cancer

目的应用2种不同方法检测晚期非小细胞肺癌（NSCLC）患者肿瘤组织和恶性胸腔积液中表皮生长因子受体（EGFR）的突变率，探讨其与临床病理学特征的关系，并比较2种检测方法的一致率。方法收集晚期NSCLC患者经甲醛固定的石蜡包埋肿瘤组织241例和其中14例的配对恶性胸腔积液，用蝎型探针扩增阻遏突变系统（蝎型探针ARMS）检测EGFR基因第18、19、20、21外显子上的29种突变，并分析EGFR基因突变与患者临床病理学特征的相关性，以及用直接测序法检测其中37例肿瘤组织EGFR基因突变状态，比较2种检测方法的一致性。结果114例晚期NSCLC患者标本中检测到EGFR基因突变，突变率为48．7％（114／234）；其中19外显子缺失突变65例，21外显子点突变L858R 39例，二者占所有突变类型的91．2％（104／114）。14例肿瘤组织和恶性胸腔积液配对的标本检测结果完全一致。应用ARMS和直接测序两种方法检测37例肿瘤组织标本的一致率为94．6％。另外，女性患者突变率（55．9％）高于男性患者（42．2％），差异有统计学意义（P〈0．05）；而不同吸烟史、组织分级以及病理类型的晚期NSCLC患者EGFR基因突变状态差异无统计学意义。结论晚期NSCLC患者EGFR基因突变率高；用ARMS法检测EGFR基因突变状态具有较高的敏感度，适用于靶向治疗患者的筛选。

Objective To investigate the mutations of epidermal growth factor receptor （EGFR） in tumor tissue and pleural effusion in advanced non-small cell lung cancer （NSCLC） patients, and to analyze the relationship between EGFR mutations and the clinicopathologic characteristics. Methods Two-hundred and forty-one cases of formalin-fixed, paraffin-embedded tumor tissues and 14 paired pleural effusions from advanced NSCLC patients were collected. Twenty-nine different EGFR mutations in exons 18-21 were assessed by scorpions and amplification refractory mutation system （scorpions ARMS） using real time PCR. The relationship between the EGFR mutations and clinical parameters was analyzed using statistical methods. EGFR mutation of 37 cases were detected with direct sequencing, and assessed the sensitivity, the specificity and the accuracy of scorpions ARMS. Results EGFR somatic mutations were detected in 114 of 234 advanced NSCLC patients, with the mutation rate of 48.7%, including deletions in exon 19 in 65 patients and point mutation of L858R in exon 21 in 39 patients ; both accounting for 91.2% （104/114） of all types of EGFR mutations. The test results of 14 paired pleural effusion specimens were entirely the same to the tissues. The concordance rate of 2 different detection methods was 94. 6%. Mutation rate was higher in women （55.9%） than in men （42. 2% ） , and there was no difference in mutation rates between smokers and non-smokers; patients in stage m B and stage IV; adenocarcinoma and non-adenocarcinoma. Conclusions EGFR somatic mutations appear to occur frequently in Chinese. Scorpions ARMS technology is a sensitive method to detect EGFR mutations and is suitable for screening patients who would likely respond to EGFR inhibitors therapy.