DNA直接测序技术检测EGFR基因及突变分析

目的：探讨非小细胞肺癌（NSCI，C）患者表皮生长因子受体（EGFR）基因突变的情况及EGFR-TKI的治疗效果。方法：从94例晚期NSCLC患者的肿瘤组织中提取DNA，采用DNA直接测序技术检测EGFR基因18、19、20、21外显子的突变情况。对EGFR基因突变可评估患者采用TKI口服治疗（吉非替尼250mg／d、厄洛替尼150mg／d），评价客观有效率（RR）、疾病控制率（DCR）及无疾病进展时间。结果：94例患者中39例（41．5％）存在EGFR基因突变，其中27例外显子19突变．2例外显子20突变，9例外显子21突变、1例外显子18突变。女性患者突变率显著高于男性（56．1％vs30．2％．P〈0．05）；腺癌患者突变率高于非腺癌患者（48％／）815．8％，P〈0．05）；不吸烟者突变率显著高于吸烟者（50．8％vs20．7％，P〈0．05）。16例患者口服TKI后部分缓解5例、疾病稳定7例、疾病进展PD4例，客观有效率为31．2％，疾病控制率为75．0％，截至随访结束，仍有14例患者生存，无疾病进展时间为（11．3l±4．44）个月，1年生存率为43．8％，、结论：DNA直接测序检测晚期NSCLC患者EGFR基因突变具有高度敏感性，以EGFR基因突变结果为依据，应用TKI治疗晚期NSCL C疗效明显。

Direct gene sequencing for measurement of epithelial growth factor receptor mutation

Objective：To determine the epithelial growth factor receptor （EGFR） mutation and the effects of EGFR tyrosine kinase inhibitor （EGFR-TRI） in patients with non-small cell lung carcinoma （NSCLC）. Methods ： The DNA was extracted from 94 patients with NSCLC to detect exon 18-21 mutations of EGFR gene via direct DNA sequencing. 16 patients with EGFR mutation received EGFR-TKI therapy （gefitinib 250mg/d plus erlotinib 150mg/d orally） for assessment of the objective effective rate （RR） , disease control rate （DCR） and progression-free survival （PFS）. Results： Of 94 patients analyzed, 39 （41.5%） had EGFR gene mutation, in whom 27 cases had exon 19 mutation, 2 had exon 20 mutation, 9 had exon 21 mutation and a single case had exon 18 mutation. A higher mutation rate was found in females compared with males （56.1% vs. 30. 2% , P 〈 0.05） , in patients with adenoma compared with those who had non-adenoma （48.0% vs. 15.8%, P 〈 0.05 ） and in never-smokers compared with ever-smokers （50.8% vs. 20.7% , P 〈 0.05）. Of the 16 patients receiving EGFR-TKI, 5 had partial remission, 7 retained stability and 4 showed progression, corresponding to an objective response and disease control rate of 31.2% and 75% , respectively. As of the end of follow-up, there were 14 cases of survival who yielded the progression-free survival of （11.31± 4.44） months and the 1-year survival of 43.8%. Conclusion： Direct DNA sequencing is a sensitive method to detect EGFR gene mutation. TKI is effective for the treatment of advanced NSCLC as guided by EGFR gene mutation.