实时荧光定量PCR法检测非小细胞肺癌EGFR基因突变的总结分析

目的:采用实时荧光定量PCR法检测临床送检的非小细胞肺癌(NSCLC)标本中表皮生长因子受体(EGFR)基因的突变情况,分析其突变率、突变特征及其与临床病理间的相关性。方法:将101例石蜡包埋NSCLC标本行显微切割后抽提DNA,采用实时荧光定量PCR法检测EGFR基因酪氨酸激酶编码区第19和21号外显子的突变。结果:101例NSCLC标本中共检出EGFR基因第19号、第21号外显子突变21例(20.8%),突变率分别为61.9%(13/21)和38.1%(8/21),其又以19 del-E746-A750和21 L858R为主要突变类型,发生率分别为42.9%(9/21)和38.1%(8/21)。EGFR基因总突变率与年龄无关(P>0.05);但女性患者中EGFR基因突变率(27.3%,12/44)高于男性患者(15.8%,9/57;P

Detection of EGFR mutation in non-small cell lung cancer by real-time PCR and its correlation with clinico.pathological characteristics

Objective： To analyze the frequency of epidermal growth factor receptor （EGFR） gene mutation and char- acteristics in non-small cell lung cancer （NSCLC） samples by real-time PCR. Methods： Clinical samples from 101 NSCLC patients were prepared for EGFR gene mutation analysis, of them 40 were surgical specimens （including 8 metastatic tumors）, 61 were biopsy or needle aspiration specimens. All samples were processed from paraffin embedded blocks and DNA was extracted after mierodissection. Real-time PCR was used to detect EGFR mutation at tyrosine kinase domain coding region involving exon 19 and 21. Results： （~） Of the 101 samples, mutations were detected in 21 cases （20.8%）. The mutation rates of exon 19 and 21 were 61.9%（13/21） and 38.1%（8/21）, respectively; the main mutations were 19 del-E746-A750 and 21 L858R, and the detection rates were 42.9% （9/21） and 38.1%（8/21）, respectively. （~） There was no significant correlation of EGFR mutation with age. （~ EGFR mutation rate was higher in female patients （27.3%,12/44） than in male patients （15.8% ,9/57; P〈0.01）. （~） All of the mutations were detected in adenocarcinomas and adenosqua- mous carcinomas, the mutation rate was 28.8% （21/73）. None was found in squamous cell carcinomas （13） and large cell carcinoma or poorly differentiated carcinomas （15）. （~） Biopsy or needle aspiration specimens showed much lower EGFR mutation detection rate （14.8%, 9/61 ） than that of surgical samples （30.0%, 12/40; P〈0.OI ）. Conclusions： EGFR muta- tions are more frequent in female patients and in patients with adenocarcinoma. The main types of mutation are 19 del- E746-A750 and 2l L858R. Detection rate of EGFR mutation may be influenced by different types of sample, false nega- tive may happen in biopsy and needle aspiration specimens.