Next-Generation Sequencing of Liquid-Based Cytology Non-Small Cell Lung Cancer Samples
Next-Generation Sequencing of Liquid-Based Cytology Non-Small Cell Lung Cancer Samples
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DOI：
10.1002/cncy.21812
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发表时间：
2017-03-01
影响因子：
3.4
通讯作者：
Cheng, Yu-Wei
中科院分区：
文献类型：
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作者：
Reynolds, Jordan P.;Zhou, Yaolin;Cheng, Yu-Wei
背景：在非小细胞肺癌（NSCLC）中，利用液体细胞学（LBC）样本（如内镜超声引导下的细针穿刺）残留的细胞颗粒检测突变的表皮生长因子受体（EGFR）已经通过等位基因特异性聚合酶链反应得到验证。本研究的目的是验证下一代测序（NGS）技术用于检测LBC残留细胞颗粒的基因突变。方法：从保存在PreservCyt中已知EGFR突变状态的腺癌LBC样本中提取存档DNA。使用Ion AmpliSeq Cancer Hotspot Panel v2化学试剂和OneTouch 2仪器对基因组DNA进行多重扩增和富集；随后在离子个人基因组机平台上进行半导体测序。利用NextGENe和Torrent Suite生物信息学工具分析6个nsclc相关基因（BRAF、EGFR、ERBB2、KRAS、MET和磷脂酰肌醇-4,5-二磷酸3-激酶催化亚基a [PIK3CA]）的突变热点。结果：常见的EGFR序列变化，包括4个L858R突变，3个外显子19缺失和1个外显子20插入，在检测平台之间的一致性为100%。在ERBB2、KRAS、MET和PIK3CA基因的突变热点也发现了不太常见的NSCLC变异。结论：使用NGS可以成功地对LBC样本的残留细胞颗粒进行NSCLC突变分析。这种方法可以同时检查多个突变热点，及时改善患者护理。(C) 2017美国癌症协会。
BACKGROUND: The detection of mutated epidermal growth factor receptor (EGFR) in non-small cell lung cancer (NSCLC) with residual cell pellets derived from liquid-based cytology (LBC) samples (eg, endoscopic ultrasound-guided fine-needle aspiration) has been validated with allele-specific polymerase chain reaction. The aim of this study was to validate next-generation sequencing (NGS) technology for detecting gene mutations with residual cell pellets from LBC. METHODS: Archived DNA extracted from LBC samples of adenocarcinoma stored in PreservCyt with a known EGFR mutation status was retrieved. Genomic DNA was multiplex-amplified and enriched with Ion AmpliSeq Cancer Hotspot Panel v2 chemistry and the OneTouch 2 instrument; this was followed by semiconductor sequencing on the Ion Personal Genome Machine platform. The mutation hotspots of 6 NSCLC-related genes (BRAF, EGFR, ERBB2, KRAS, MET, and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit a [PIK3CA]) were analyzed with NextGENe and Torrent Suite bioinformatics tools. RESULTS: The commonly identified EGFR sequence changes, including 4 L858R mutations, 3 exon 19 deletions, and 1 exon 20 insertion, were in 100% concordance between the assay platforms. Less common NSCLC variants were also found in the mutation hotspots of ERBB2, KRAS, MET, and PIK3CA genes. CONCLUSIONS: NSCLC mutation analysis using NGS can be successfully performed on residual cell pellets derived from LBC samples. This approach allows the simultaneous examination of multiple mutation hotspots in a timely manner to improve patient care. (C) 2017 American Cancer Society.