应用高通量测序技术检测甲状腺乳头状癌相关基因变异

Application and analysis of NGS based detection in thyroid papillary carcinoma

目的探讨高通量测序技术(next generation sequencing,NGS)在甲状腺乳头状癌(papillary thyroid cancer,PTC)相关的15个靶基因变异检测中的应用价值。方法收集2019年1月至2020年6月就诊的188例PTC患者手术切除的癌组织石蜡包埋标本,采用NGS技术检测其中15个目标基因;利用GATK 4.0.2.0、VarScan.v2.3.9和Factera 1.4.4软件分析基因变异的特点。结果共计80.32%(151/188)的标本检出突变。其中74.47%(140/188)的标本检出1个点突变或基因融合突变,5.32%(10/188)的标本同时检出2个点突变,0.53%(1/188)的标本同时检出3个点突变。151例突变标本中共检出点突变及基因融合突变163个,其中BRAF基因检出率最高。在所有双突变的标本中,2个不同突变等位基因突变丰度基本一致。在3个基因同时发生突变的标本中,TERT和NRAS基因等位基因突变丰度基本一致,而TP53基因等位基因突变丰度显著低于其他2个基因。PTC患者基因突变与性别(χ^2=0.585,P>0.05)、年龄(χ^2=0.575,P>0.05)及临床分期(χ^2=0.711,P>0.05)均无关。结论NGS技术检测PTC相关基因突变位点具有经济、效率高和可定量的优势,可为患者的诊断、预后和个体化治疗提供依据。

Objectives To explore the application of next-generation sequencing(NGS)based detection for a panel of 15 target genes mutations and analyzed the gene spectrum in papillary thyroid carcinoma(PTC).Methods A total of 188 FFPE(formalin-fixed,paraffin-embedded)tissue specimens from surgically removed PTC samples were detected with 15 target genes by NGS.Gatk 4.0.2.0,varscan V2.3.9 and Factera 1.4.4 software were used to analyze the characteristics of gene variation.Results The gene mutations of NGS were detected in 80.32%of all samples(151/188).One point mutation or gene fusion mutation were carried in single gene of 74.47%(140/188)samples,two point mutations were simultaneously harboured in 5.32%(10/188)samples and triple different gene point mutations were carried in 0.53%(1/188)sample.A total of 163 point mutations and gene fusion mutations were detected in 151 mutation samples.The BRAF mutation was the most common mutation type in PTC patients.For the samples carried two mutations,the allelic frequency of mutations was similar.However,for the samples harboured three different gene mutations,the allelic frequency of NRAS mutation was similar to TERT mutation but TP53 mutation showed much lower abundance.There was no correlation between gene mutation and gender(χ^2=0.585,P>0.05),age(χ^2=0.575,P>0.05)and clinical stages(χ^2=0.711,P>0.05)in PTC patients.Conclusion NGS-based detection method may comprehensively detect the mutations in PTC related genes,and provide the basis for diagnosis,prognosis and individualized treatment of patients.