肺癌患者呼出气冷凝液与血液中EGFR、KRAS基因及其变异的检测

Detection of exhaled breath condensate and blood EGFR and KRAS genes and their variation in lung cancer patients

目的:研究肺癌患者检测呼出气冷凝液(EBC)中表皮生长因子受体(EGFR)、KRAS基因及其变异的可行性,与肺癌患者血液中EGFR、KRAS基因及其变异的检测进行对比研究,并结合病理及影像学等临床资料分析其临床意义。方法:将收治的30例肺癌患者作为研究组,选取同一时间健康志愿者30例作为对照组。分别采集两组研究对象的EBC和血液标本,检测EBC和血液EGFR、KRAS基因表达及变异性情况。结果:研究组和对照组对象EBC标本中,通过多种检测方法均未提取到EBC样本基因组DNA,无法扩增出检测目的基因。研究组中EGFR基因中有18例发生突变,其中有15例样本中19外显子缺失敏感型突变,仅3例发现21外显子L85 8R突变,KRAS基因仅1例发生突变,为Gly12Asp突变。15例19外显子缺失敏感型突变中腺癌12例,鳞癌3例。对照组中EGFR基因中均未发生突变。KRAS基因中均未发生突变。研究组EGFR基因野生型和突变型比例与对照组比较,差异有统计学意义(P<0.05);KRAS基因野生型和突变型两组差异无统计学意义(P>0.05)。结论:用EBC标本检测肺癌患者中EGFR、KRAS基因是一个理想的无创的方法,但目前因多种原因及条件限制无法完成。血液中EGFR、KRAS基因及其变异性是目前比较实用的方法,创伤小。

Objective To study the feasibility of detecting EGFR and KRAS genes and their variants in exhaled breath condensate(EBC)of lung cancer patients,and compare them with the detection of EGFR and KRAS genes and their variants in blood of lung cancer pat ients.Method 30 lung canc er patients were selected as the study group,and 30 healthy volunteers at the same time from the physical examination center of our hospital were selected as the control group.EGFR and KRAS gene expression and variability were detected in exhaled breath condensate and blood samples of the two groups.Results The genomic DNA of the respiratory condensate sample was not extracted from the EBC sam ples of the study group and the control group by various detection methods,and the target gene could not be amplified.There were 18 mutations in EGFR gene in the study group,of which 19 mutations were sensitive to exon deletion in 15 samples,only 3 mutations in exon 21 L858R.Only 1 mutation occurred in the KRAS gene,which was a Gly12Asp mutation.In 15 cases,19 exon deletion sensitive mutations were found in 12 cases of adenocarcinoma and 3 cases of squamous cell carcinoma.In the control group,no mutation was found in the EGFR gene.No mutation was found in the KRAS gene.The wild type and mutation ratio of EGFR gene in the study group were significantly different from those in the control group(P<0.05);There was no difference in the wild-type and mu tant KRAS between the two groups(P>0.05).Conclusion It is an ideal noninvasive method to detect EGFR and K RAS genes in lung cancer patients with exhaled air condensate,but it can not be completed due to various reasons and conditional limitations.EGFR and KRAS genes in blood and their variability are relatively practical methods with little trauma.