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中国癌症防治杂志

中国癌症防治杂志 ›› 2019, Vol. 11 ›› Issue (6): 480-484.doi: 0.3969/j.issn.1674-5671.2019.06.06

• 基础研究 • 上一篇    下一篇

基于靶向二代测序技术检测肝细胞癌线粒体DNA突变

  

  1. 内蒙古医科大学基础医学院；空军军医大学基础医学院生理教研室；空军军医大学第一附属医院肝胆外科

  • 出版日期:2019-12-25 发布日期:2020-01-19
  • 通讯作者: 郭旭 E-mail：lfmmuxguo@163.com
  • 基金资助:
    国家自然科学基金项目（81602463）；陕西省科技计划项目（2018ZDXM-SF-061）

Detection of mitochondrial DNA mutations in hepatocellular carcinoma by targeted next generation sequencing

  • Online:2019-12-25 Published:2020-01-19

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摘要： 目的 探讨肝细胞癌（hepatocellular carcinoma，HCC）患者不同类型样本的线粒体DNA（mitochondrial DNA， mtDNA）突变情况。方法 收集2017年9月23日至2017年12月18日在空军军医大学第一附属医院经病理确诊的5例HCC患者的癌组织及癌旁组织(距癌组织≤3 cm)、外周血细胞及血浆样本，通过全基因组测序数据分析癌组织及血浆样本的mtDNA拷贝数，并基于靶向二代测序（next generation sequencing，NGS）技术分析HCC患者癌组织、癌旁组织、外周血细胞及血浆中mtDNA突变的情况。结果 全基因组测序结果显示，与癌组织相比，血浆中的mtDNA拷贝数显著降低（Z＝-2.023，P＝0.008）。靶向捕获测序结果显示，癌组织、癌旁组织、外周血细胞、血浆样本可分别鉴定出181、180、181、179个同质性突变和33、42、3、78个异质性突变。癌组织、癌旁组织、血浆中可分别检测到10、39、56个特异性突变位点，而血浆中的mtDNA突变频率与癌组织比较差异无统计学意义（P=0.219）。结论 基于靶向NGS技术在肝细胞癌患者血浆中可检测到肿瘤来源的mtDNA突变，且能够准确检测出血浆中的低频突变，可作为组织活检的有力补充。

关键词: 肝细胞癌, 二代测序, 线粒体DNA, 体细胞突变

Abstract: Objective To investigate mitochondrial DNA（mtDNA） mutations between different types of samples in hepatocellular carcinoma （HCC） patients. Methods The tumor tissues，adjacent tissues （≤3 cm from the tumor tissue），peripheral blood cells，and plasma of 5 patients with HCC diagnosed pathologically in the First Affiliated Hospital of Air Force Medical University were collected from September 23，2017 to December 18，2017. mtDNA copy number of tumor tissues and plasma samples were detected by whole genome sequencing. mtDNA mutations in tumor tissues，adjacent tissues，peripheral blood cells and plasma samples were analyzed by targeted next generation sequencing（NGS）. Results Whole genome sequencing results showed that the mtDNA copy number in plasma was significantly reduced compared with tumor tissues（Z＝-2.023，P＝0.008）. Targeted capture sequencing results showed that 181，180，181，179 homogeneous mutations and 33，42，3，78 heterogeneous mutations were identified in tumor tissues，adjacent tissues，peripheral blood cells and plasma samples，respectively. 10，39，and 56 specific mutation sites were detected in tumor tissues，adjacent tissues，and plasma，respectively. The frequency of mtDNA mutations in plasma was not significantly lower than that in tumor tissues（P=0.219）. Conclusion  Targeted next generation sequencing can detect mtDNA mutations in plasma samples of patients with hepatocellular carcinoma，and accurately discover the low-frequency mutations in plasma，which can be a powerful complement to tissue biopsy.

Key words: Hepatocellular carcinoma, Next generation sequencing, Mitochondrial DNA, Somatic mutation

中图分类号: 

  • R735.7