二代测序和免疫组织化学在子宫内膜癌分子分型中的比较  

Comparative study of next generation sequencing and immunohistochemistry on molecular classification of endometrial carcinoma

作　　者：陈婷婷 陶祥 刘天齐 周先荣 Chen Tingting;Tao Xiang;Liu Tianqi;Zhou Xianrong(Department of Pathology,Obstetrics and Gynecology Hospital of Fudan University,Shanghai 200011,China)

机构地区：[1]复旦大学附属妇产科医院病理科,上海200011

出　　处：《中华病理学杂志》2023年第6期580-585,共6页Chinese Journal of Pathology

摘　　要：目的探讨子宫内膜癌不同技术方法分子分型的差异以探索符合国情的分子分型方案。方法收集复旦大学附属妇产科医院病理科2021年4月至2022年3月诊断的子宫内膜癌254例,采用二代测序和免疫组织化学检测综合方案进行分子分型,并以POLE和TP53突变频率≥20%设定为Sanger测序检测极限值模拟其结果,比较不同组合方案下分子分型结果。结果254例子宫内膜癌患者,年龄24~89岁,平均年龄51岁。包含有POLE的第9~14号外显子、TP53全外显子和微卫星不稳定(MSI)检测的二代测序panel为较好的独立解决方案,结合MSI片段分析检测和常规免疫组织化学初筛为最佳的综合解决方案,和癌症基因组图谱(TCGA)数据及近期文献比对有最佳符合率,POLE超突变型、错配修复缺陷型、TP53突变型及非特异分子特征型占比分别为11.4%(29/254)、31.5%(80/254)、22.4%(57/254)和34.6%(88/254);如改用Sanger测序进行POLE和TP53突变检测,则分别为9.1%(23/254)、31.5%(80/254)、12.9%(33/254)和46.6%(118/254),大幅度增加了非特异分子特征型的数量;如采用Sanger测序检测POLE,其余分型均用免疫组织化学检测替代,则分别为9.1%(23/254)、42.2%(92/218)、13.8%(35/254)和40.9%(105/254),将增加错配修复缺陷组的假阳性判断。结论包含MSI检测的中小型二代测序panel为分子分型的较好解决方案,Sanger测序对POLE突变检测目前可满足基本要求,但对TP53突变检测灵敏度不够,和p53免疫组织化学判断TP53突变效率相当。进一步优化含MSI检测和POLE、TP53全外显子的中小型二代测序方案是未来符合国情的最优选择。Objective To investigate the differences in molecular classification of endometrial carcinoma(EC)between various technical methods and to explore molecular classification schemes suitable for Chinese population.Methods The study used a comprehensive scheme of next generation sequencing(NGS)and immunohistochemistry for molecular classification of 254 EC cases that were obtained at Department of Pathology,Obstetrics and Gynecology Hospital of Fudan University,Shanghai,China from April 2021 to March 2022.According to the recommended threshold of Sanger sequencing which was approximate-20%variant allele fraction(VAF),NGS data were extracted to simulate the results of Sanger sequencing.Results The 254 EC patients had a mean age of 51 years(range,24 to 89 years).Combination of POLE(9-14 exons),TP53 total exons and microsatellite instability(MSI)detection was a better single scheme than NGS alone,while combination of MSI fragment analysis and conventional immunohistochemistry was the best solution and seemed best aligned with TCGA data and recent studies.POLE ultramuted type,mismatch repair defect type,TP53 mutant type and non-specific molecular characteristic type accounted for 11.4%(29/254),31.5%(80/254),22.4%(57/254)and 34.6%(88/254)of the cases,respectively.If Sanger sequencing was adopted for POLE and TP53 detection,the frequencies of these EC types were 9.1%(23/254),31.5%(80/254),12.9%(33/254)and 46.6%(118/254),respectively,with greatly increasing non-specific molecular characteristics cases.If POLE was detected by Sanger sequencing and others by immunohistochemistry,they were 9.1%(23/254),42.2%(92/218),13.8%(35/254)and 40.9%(105/254),respectively,with increasing the false positive rates of the mismatch repair defect group.Conclusions Small and medium-sized NGS panels with MSI detection is a better solution than NGS alone.Sanger sequencing is currently available for POLE mutation detection,which is not sensitive enough for TP53 mutation detection,and seems equivalent to the efficiency of TP53 by immunohistochemist

关 键 词：子宫内膜肿瘤 基因 P53 微卫星不稳定性 

分 类 号：R737.33[医药卫生—肿瘤]