46例前列腺癌患者的组织DNA及血液ctDNA测序分析  

Sequencing analysis on tissue DNA and blood ctDNA from 46 patients with prostate cancer

作　　者：唐堂 唐朋 谭信韬 冉强 彭松 刘秋礼 徐静 江军[1] TANG Tang;TANG Peng;TAN Xintao;RAN Qiang;PENG Song;LIU Qiuli;XU Jing;JIANG Jun(Department of Urology,Daping Hospital,Army Medical University(Third Military Medical University),Chongqing,400042,China)

机构地区：[1]陆军军医大学(第三军医大学)大坪医院泌尿外科,重庆400042

出　　处：《第三军医大学学报》2021年第17期1658-1666,F0003,共10页Journal of Third Military Medical University

基　　金：国家自然科学基金青年科学基金项目(81802558);陆军军医大学科研项目(2017XYY07)。

摘　　要：目的探讨前列腺癌患者组织DNA与血液ctDNA检出的突变情况,分析基因突变与临床特点和治疗的相关性。方法采用探针杂交捕获和Illumina高通量测序为基础的靶向二代测序技术检测本院泌尿外科46例病理诊断明确的前列腺癌患者的前列腺组织标本(n=31)以及外周血液标本(n=32)循环肿瘤细胞DNA(circulating tumor DNA,ctDNA)的突变情况,检测范围包括1021个基因的全外显子区域及部分内含子区域中的4种突变类型(点突变、小片段的插入缺失、拷贝数变异和目前已知的融合基因)。结果在31例患者的前列腺组织标本中最常被检出的突变基因为TP53(29%,9/31),其次为CDK12(22.6%,7/31)、FOXA1(22.6%,7/31)与JAK1(16.1%,5/31)。在32例外周血ctDNA标本中检测出的常见突变基因为AR(34.3%,11/32)、APC(25%,8/32)、CDK12(18.8%,6/32)、DNMT3a(18.8%,6/32)与TP53(18.8%,6/32)。TMPRSS2-ERG融合基因在所有检测患者中检出率为13%(6/46)。其中1例患者同时存在TMPRSS2-ERG、VEGFA-TMPRSS2及ERG-VEGFA融合突变。46例患者中同源重组相关基因的体细胞突变检出率为21.7%,携带有胚系同源重组基因突变患者占8.7%。且携带(无论胚系或体细胞)同源重组基因突变患者的发病年龄更低、Gleason评分更高、转移灶数量更多,特别是携带胚系突变的患者更显著。结论血液ctDNA和传统的组织DNA样本均可检出基因突变;血液ctDNA检测可动态监控前列腺癌基因频谱在治疗过程中的变化,有助于及时调整治疗方案。Objective To investigate the mutations in the tissue DNA and plasma circulating tumor DNA(ctDNA)from patients with prostate cancer,and analyze their association with clinical characteristics and treatment.Methods Based on probe hybridization capture and Illumina high-throughput sequencing,a targeted second-generation sequencing was used to analyze the mutations in 31 tumor tissues and 32 peripheral blood specimens from 46 patients with pathologically diagnosed prostate cancer.The tumor tissues and peripheral blood specimens were sequenced in the whole exons area and part of the introns area of 1021 genes for 4 types of mutations(point mutation,loss/insert of small fragments,copy number variation and currently known fusion genes).Results The most frequently mutated gene was TP53(29.0%,9/31),followed by CDK12(22.6%,7/31),FOXA1(22.6%,7/31)and JAK1(16.1%,5/31)in 31 tissue samples.The most common mutant genes in the 32 blood samples were AR(34.3%,11/32),APC(25.0%,8/32),CDK12(18.8%,6/32),DNMT3a(18.8%,6/32)and TP53(18.8%,6/32).TMPRSS2-ERG fusion gene was detected in 13.0%(6/46)of the 46 patients.One patient had TMPRSS2-ERG,VEGFATMPRSS2,and ERG-VEGFA fusion mutations simultaneously.The somatic mutation of homologous recombination(HR)associated genes were found in 21.7%(10/46)of all the patients,and those with germline mutations in the HR gene accounted for 8.7%(4/46)of all the patients.Moreover,the patients with HR gene mutations(either germline or somatic),especially those with germline mutations,had younger onset-age,higher Gleason score,and more metastases.Conclusion Gene mutation can be detected from both blood ctDNA and traditional tissue DNA.Dynamic blood ctDNA testing can monitor the changes in gene spectrum of prostate cancer,and is helpful to adjust treatment scheme in time.

关 键 词：前列腺癌 二代测序 全外显子测序 基因突变 同源重组基因 

分 类 号：R394.3[医药卫生—医学遗传学] R730.43[医药卫生—基础医学]