非小细胞肺癌患者细胞学标本EGFR基因突变检测及其临床病理意义

Detection of EGFR gene mutation in non-small cell lung cancer and its clinical significance

目的:探讨细胞学标本在非小细胞肺癌(NSCLC)的诊断及个体化治疗中的临床应用价值。方法:收集352例新鲜细胞学标本制片后,行常规HE染色;同时选择TTF-1、NapsinA、CK7、CEA、CD56、Syn、P63、CK5/6、WT-1、E-cadherin等抗体对来源不明的肿瘤细胞进行免疫细胞化学标记,并对明确诊断为NSCLC的病例,采用突变扩增阻滞系统(ARMS)检测表皮生长因子受体(EGFR)基因突变情况。结果:352例患者中,345例有癌细胞。经临床及免疫细胞化学证实345例恶性细胞中,NSCLC有335例,且NSCLC细胞学标本中有302例DNA提取成功,占90.15%(302/335)。EGFR基因检测结果显示,EGFR共突变123例,总突变率为40.73%(123/302)。其中,第18、19、20、21外显子的突变率分别为0.99%(3/302)、19.21%(58/302)、0.66%(2/302)和19.87%(60/302);EGFR 18、19、21外显子突变占EGFR突变总数的98.37%(121/123)显著高于EGFR 20外显子突变(P<0.05)。302例患者中,女性患者EGFR突变率为54.35%(75/138),明显高于男性患者29.27%(48/164)(P<0.05);非吸烟患者EGFR的突变率为51.49%(104/202),显著高于吸烟者19%(19/100)(P<0.05)。276例腺癌中EGFR突变率44.20%(122/276);非腺癌EGFR突变率4.34%(1/23);腺癌EGFR突变率明显高于其他类型(P<0.05)。结论:利用新鲜细胞学标本,结合免疫细胞化学标记和ARMS分子病理技术有助于晚期非小细胞肺癌的诊断,并为表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)个体化治疗提供可靠依据。

OBJECTIVE：To investigate the clinical value of cytological specimens in the diagnosis and the individualized treatment of non-small cell lung cancer（NSCLC）.METHODS：Cytological specimens were stained with HE and with immunocytochemical method to detect expression of an appropriate set of antibodies（TTF-1,NapsinA,CK7,CEA,CD56,Syn,P63,CK5/6,WT-1,E-cadherin）,in tumor cells of unknown origin.The epidermal growth factor receptor（EGFR） gene was detected by the amplification refractory mutation system（ARMS） in NSCLC.RESULTS：In 352 patients,345 cases were found cancer cells on the cytological specimens.According to the clinical history and immunocytochemical staining of the 345 malignant cases,335 were NSCLC cytological samples,and the DNA of 302 NSCLC samples were extracted successfully（satisfaction rate at 90.15%）.EGFR mutations were detected in 123 of the 302 specimens（40.73%,123/302） and the frequencies of EGFR mutations in exon18,19,20（T790M）,21 were 0.99%（3/302）,19.21%（58/302）,0.66%（2/302） and 19.87%（60/302）,respectively.Higher frequencies of EGFR mutations were detected in exons 18,19 and 21（98.37%,121/123） than in exon 20（P〈0.05）.EGFR mutations were more frequently detected in women（54.35%,75/138） than in men（29.27%,48/164）（P〈0.05）,and in non-smokers（51.49%,104/202） than in smokers 19%（19/100）（P〈0.05）.Mutations were identified in 44.20%（122/276） cases of adenocarcinoma and 4.34%（1/23） in nonadenocarcinoma.Mutation of EGFR gene in adenocarcinoma was higher than that in non-adenocarcinoma（P〈0.05）.CONCLUSION：The use of cytological specimens in combination with immunocytochemical staining and molecular techniques helps in the diagnosis of advanced cancer and individualized treatment option of NSCLC.