蝎形探针扩增阻滞突变系统检测肺癌患者血清与肿瘤组织中表皮生长因子受体基因突变  

Detection of epidermal growth factor receptor gene mutations in tumor and matched serum in lung cancer patients by Scorpions amplification refractory mutation system

作　　者：周南[1] 李龙芸[1] 郑直[4] 李单青[2] 崔巍[3] 翁姗姗[1] 王孟昭[1] 王树兰[1] 钟巍[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院呼吸科,北京100730 [2]中国医学科学院北京协和医学院北京协和医院胸外科,北京100730 [3]中国医学科学院北京协和医学院北京协和医院检验科,北京100730 [4]中国医学科学院基础医学研究所北京协和医学院基础医学院,北京100005

出　　处：《癌症进展》2011年第1期19-26,共8页Oncology Progress

摘　　要：目的应用蝎形探针扩增阻滞突变系统(Scorpions ARMS)检测肺癌患者血清游离DNA和肿瘤组织中表皮生长因子受体(EGFR)基因突变,探索二者间的一致性。方法用硅胶膜离心柱法提取石蜡切片中DNA,比较硅胶膜离心柱法、酚氯仿法及磁珠法提取血清游离DNA的结果。应用Scorpions ARMS检测石蜡包埋肿瘤组织以及匹配患者血清游离DNA中EGFR基因第18、19、20和21外显子的突变。结果 35例肺癌组织标本和匹配的血清经上述方法测定,其中EGFR基因总突变率分别为54.3%和17.1%,敏感突变分别为31.4%和8.6%,血清和肿瘤组织EGFR基因突变的一致率为60%,Ⅲ和Ⅳ期患者血清EGFR基因突变率(35.7%)显著高于Ⅰ和Ⅱ期患者(4.5%)。结论血清游离DNA可能为监测Ⅲ和Ⅳ期肺癌患者EGFR基因突变提供新的来源;Scorpions ARMS是检测血清游离DNA和原发肿瘤组织中EGFR基因突变的可靠有效方法;磁珠法是提取血清DNA的有效方法,值得进一步探讨。Objective To detect the epidermal growth factor receptor （EGFR） mutations in tumor and matched serum in lung cancer patients by Scorpions Amplification Refractory Mutation System （ Scorpions ARMS）. Methods Spin column method was used to extract DNA from formalin-fixed paraffin-embedded resected tumors find three methods （ spin column, magnetic beads, and phenol chloroform） were used to extract cell-free DNA from serum. Thirty five tumor sam-ples and matched serum specimens from patients with lung cancer were analyzed for EGFR mutations in exons 18, 19, 20, and 21 by Scorpions ARMS. Results EGFR mutations were detected in 54. 3% （19/35） of the tumor samples and 17. 1% （6/35） of the serum specimens. EGFR mutation status in the tumors and serum samples was concordant in 21 （60%） of the 35 pairs. EGFR mutation rate in the serum specimens of stage mand 1V patients （35.7%, 5/14） was significantly higher than in stage Ⅰ and Ⅱ patients （4. 5% , 1/21 ）. Conclusion Cell-free DNA in serum offers the possibility of monitoring EGFR mutations in primary tumors in patients with stage Ⅲ and IV lung cancer. Scorpions ARMS is reliable and effective for the detection of EGFR nmtations. Magnetic beads method is more effective than the other two methods for extracting DNA from serum.

关 键 词：肺癌 表皮生长因子受体基因突变 

分 类 号：R734.2[医药卫生—肿瘤]