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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 mutations of epidermal growth factor receptor (EGFR) gene in serum free DNA and tumor tissue of patients with lung cancer by Scorpions ARMS (Scorpions ARMS), and to explore the consistency between them. Methods The DNA in paraffin sections was extracted by silica gel column centrifugation, and the results of serum free DNA extraction by silica gel column centrifugation, phenol-chloroform method and magnetic bead method were compared. Scorpions ARMS was used to detect mutations in exons 18, 19, 20, and 21 of the EGFR gene in paraffin-embedded tumor tissue and matched serum free DNA. Results 35 cases of lung cancer tissue samples and matched serum were determined by the above method. The total mutation rates of EGFR gene were 54.3% and 17.1% respectively, the sensitive mutations were 31.4% and 8.6% respectively. The consistent rates of EGFR gene mutations in serum and tumor tissues were The rate of EGFR gene mutation (35.7%) was significantly higher in 60%, Ⅲ and Ⅳ patients than in Ⅰ and Ⅱ patients (4.5%). Conclusion Serum free DNA may provide a new source for monitoring the EGFR gene mutation in stage Ⅲ and Ⅳ lung cancer patients. Scorpions ARMS is a reliable and effective method to detect EGFR gene mutation in serum free DNA and primary tumor tissue. Effective method, it is worth further study.