基于下一代测序的胃癌和结直肠癌患者粪便中的基因突变。
Gene mutations in stool from gastric and colorectal neoplasia patients by next-generation sequencing.
机构信息
Department of Pathology, Faculty of Medicine, University of Helsinki, Helsinki 00014, Finland.
Department of Pathology, University of Helsinki and HUSLAB, Helsinki University Hospital, Helsinki 00014, Finland.
出版信息
World J Gastroenterol. 2017 Dec 21;23(47):8291-8299. doi: 10.3748/wjg.v23.i47.8291.
AIM
To study cancer hotspot mutations by next-generation sequencing (NGS) in stool DNA from patients with different gastrointestinal tract (GIT) neoplasms.
METHODS
Stool samples were collected from 87 Finnish patients diagnosed with various gastric and colorectal neoplasms, including benign tumors, and from 14 healthy controls. DNA was isolated from stools by using the PSP Spin Stool DNA Plus Kit. For each sample, 20 ng of DNA was used to construct sequencing libraries using the Ion AmpliSeq Cancer Hotspot Panel v2 or Ion AmpliSeq Colon and Lung Cancer panel v2. Sequencing was performed on Ion PGM. Torrent Suite Software v.5.2.2 was used for variant calling and data analysis.
RESULTS
NGS was successful in assaying 72 GIT samples and 13 healthy controls, with success rates of the assay being 78% for stomach neoplasia and 87% for colorectal tumors. In stool specimens from patients with gastric neoplasia, five hotspot mutations were found in , and genes, in addition to seven novel mutations. From colorectal patients, 20 mutations were detected in , , , , , , , , , and . Healthy controls did not exhibit any hotspot mutations, except for two novel ones. and were the most frequently mutated genes in colorectal neoplasms, with five mutations, followed by with two mutations. was the most commonly mutated gene in stools of patients with premalignant/benign GIT lesions.
CONCLUSION
Our results show that in addition to colorectal neoplasms, mutations can also be assayed from stool specimens of patients with gastric neoplasms.
目的
通过下一代测序（NGS）研究来自不同胃肠道（GIT）肿瘤患者粪便 DNA 中的癌症热点突变。
方法
收集 87 名芬兰患者的粪便样本，这些患者被诊断患有各种胃和结直肠肿瘤，包括良性肿瘤，以及 14 名健康对照者。使用 PSP Spin Stool DNA Plus Kit 从粪便中提取 DNA。对于每个样本，使用 20ng 的 DNA 采用 Ion AmpliSeq Cancer Hotspot Panel v2 或 Ion AmpliSeq Colon and Lung Cancer panel v2 构建测序文库。在 Ion PGM 上进行测序。使用 Torrent Suite Software v.5.2.2 进行变异调用和数据分析。
结果
NGS 成功检测了 72 个 GIT 样本和 13 个健康对照者，胃肿瘤的检测成功率为 78%，结直肠肿瘤的检测成功率为 87%。在胃肿瘤患者的粪便标本中，除了 7 个新突变外，还在 、 和 基因中发现了 5 个热点突变。从结直肠患者中，在 、 、 、 、 、 、 、 、 和 基因中检测到 20 个突变。健康对照者除了发现 2 个新突变外，没有显示任何热点突变。在结直肠肿瘤中， 、 、 、 、 、 、 、 、 和 基因最常发生突变，有 5 个突变，其次是 有 2 个突变。在具有癌前/良性 GIT 病变的患者的粪便中， 是最常发生突变的基因。
结论
我们的研究结果表明，除了结直肠肿瘤外，还可以从胃肿瘤患者的粪便标本中检测到突变。
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