基于分子标签二代测序技术的非小细胞肺癌驱动基因变异分析  

Analysis of driving gene mutations in non-small cell lung cancer using UMIbased next-generation sequencing

作　　者：刘小云 吴小延[1] 邵琼[1] 龙亚康 王海云 邓玲[1] LIU Xiaoyun;WU Xiaoyan;SHAO Qiong;LONG Yakang;WANG Haiyun;DENG Ling(State Key Laboratory of Oncology in South China/Collaborative Innovation Center for Cancer Medicine,Sun Yat sen University Cancer Center,Guangzhou,Guangdong,China,510060)

机构地区：[1]中山大学肿瘤防治中心华南肿瘤学国家重点实验室肿瘤医学协同创新中心

出　　处：《分子诊断与治疗杂志》2019年第6期468-473,共6页Journal of Molecular Diagnostics and Therapy

基　　金：国家自然科学基金青年基金(81602468)

摘　　要：目的 通过对非小细胞肺癌(NSCLC)患者的血浆 ctDNA标本采用基于分子标签(UMI) 的二代测序(NGS)技术进行相关驱动基因突变分析,评估血浆 ctDNA二代测序技术指导靶向用药的价 值及注意事项。 方法 纳入了163例2017年7月至2019年7月期间在中山大学附属肿瘤医院就诊的 NSCLC患者血液标本,采用基于分子标签的二代测序方法,检测患者血浆ctDNA中与NSCLC相关的 11个驱动基因的突变情况。 结果 98例初诊初治的NSCLC患者中,37.8%(37例)患者未检出肺癌驱 动基因突变。其余患者中 TP53、EGFR、ERBB2、RET、KRAS、ALK、PIK3CA、BRAF、MET、ROS1、NRAS的突 变频率分别为33%、24%、10%、8%、7%、6%、4%、3%、3%、3%和1%。57例EGFR TKI靶向治疗后进展的 NSCLC患者中,21例(36.8%)检出T790M突变,3例(27.3%)接受三代EGFR TKI治疗后进展的患者出 现T790M与T797S顺式耐药突变。其他耐药突变包括KRAS基因突变(2例)、MET基因扩增(1例)以 及PIK3CA基因突变(8例)。29例组织样本ARMS PCR检测结果阳性的患者,组织EGFR突变阳性患者 与ctDNA检测EGFR阳性患者之间的一致性为13.8%。 结论 该方法不仅能够检出初诊初治NSCLC 患者中靶向治疗的敏感突变,还可以对靶向治疗进展后的耐药突变情况进行监测。因此,可选择基于分 子标签的二代测序检测技术来指导 NSCLC患者的个性化用药。Objective Next-generation sequencing(NGS)based on unique molecular identifiers(UMI)was performed on circulating tumor DNA from non-small cell lung cancer patients to investigate its clinical application value. Methods The blood samples of 163 NSCLC patients who visited the Affiliated Cancer Hospital of Sun Yat sen University from July 2017 to July 2019 were enrolled. A panel of 150 genes was used to detect the genetic mutations of the plasma samples,the mutations of 11 driving genes related to NSCLC in plasma ctDNA of patients were detected by using the NGS based on UMI. Results Among 98 patients with TKI-nave patients(including 73 lung adenocarcinomas,8 squamous carcinomas and 17 with unknown pathological types)of NSCLC,37.8%(37 cases)had no lung cancer driving gene mutation. The mutation frequencies of TP53,EGFR,erbB2,RET,KRAS,ALK,PIK3 CA,BRAF,MET,ROS1 and NRAS were 33%,24%,10%,8%,7%,6%,4%,3%,3%,3% and 1%,respectively. Among 57 NSCLC patients who developed after EGFR-TKI targeted therapy,T790 M mutation was detected in 21(36.8%)patients,and T790 M and t797 s CIS resistant mutations were detected in 3(27.3%) patients who developed after three generations of EGFR-TKI treatment. Other resistant mutations included KRAS gene mutation(2 cases),MET gene amplification(1 case)and PIK3 CA gene mutation(8 cases). Among 29 NSCLC patients with positive EGFR mutations in tissues,only 4 matched plasma samples were detected positive,and the consistency of EGFR detection rates between matched tissue and plasma samples were 13.8%. Conclusion This method can detect the sensitive mutations of targeted therapy in patients with primary NSCLC and monitor the drugresistant mutations after the progress of targeted therapy. Therefore,the NGS based on molecular tags can provides valuable information about targeted treatment to guide the personalized medication of NSCLC patients.

关 键 词：非小细胞肺癌 二代测序 分子标签 液体活检 循环肿瘤游离DNA 耐药突变 

分 类 号：R73[医药卫生—肿瘤]