Sanger法检测分析非小细胞肺癌患者组织EGFR

目的探讨Sanger法检测非小细胞肺癌（non-small cell lung cancer，NSCLC）患者组织表皮生长因子受体（epidermal growth factor receptor，EGFR）基因突变情况，为肺癌病人“个体化靶向分子治疗”提供依据。方法收集63例NSCLC患者组织标本（石蜡切片），提取组织DNA，用EGFR外显子18、19、20和21四位点的分子扩增（PCR）试剂盒进行外显子扩增，扩增产物先后进行电泳鉴定、酶解，酶解产物PCR扩增、纯化，纯化产物用ABI-3130型基因分析仪检测得出EGFR外显子18、19、20、21的野生或突变状态结果，并分析EGFR基因突变和患者临床病理生理特征、临床靶向用药疗效的关系。结果测试显示肺腺癌患者EGFR基因19、21外显子的突变率为33．3％（21／63），而EGFR基因的突变与患者的病理组织学类型、吸烟状态有关（P〈0．05）。同时EGFR突变患者能从临床靶向用药获益。结论Sanger法检测NSCLC突变情况可为临床靶向用药提供技术支持。

Sanger method for detecting tissue EGFR in patients with non-small cell lung cancer

Objective To provide support for personalized targeted molecular therapy of lung cancer patients by exploring gene mutations of tissue epidermal growth factor receptor ( EGFR ) in nonsmall cell lung cancer.Methods Tissue samples were collected from 63 patients to extract DNA.Amplication of EGFR exons 18,19,20,and 21 was performed with PCR kit.The products of amplication were identified by dectrophoresis and then digested by enzymes.The products of digestion were amplified by PCR and then were purified.The end products were detected by the AB13130 genetic analyzer to obtain the wild-types or mutant status of EGFR exons 18,19,20,and 21.The relationship of EGFR gene mutations with the clinical pathophysiologieal characteristics and the efficacy of targeted therapy was analyzed.Results The mutation rate of EGFR exons 19 and 21 was 33.3％ ( 21/63 ),and the mutations were associated with the histological types and smoking status ( P＜ 0.05 ).The targeted therapy was beneficial for the patiens with EGFB mutation.Conclusions Sanger method for detecting gene mutations in NSCLC can provide technical support for the targeted therapy.