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非子宫内膜样卵巢上皮性肿瘤中的微卫星不稳定性:400 例病例比较免疫组化、PCR 和基于 NGS 的检测与 MMR 基因突变状态的研究。

Microsatellite instability in non-endometrioid ovarian epithelial tumors: a study of 400 cases comparing immunohistochemistry, PCR, and NGS based testing with mutation status of MMR genes.

机构信息

Department of Pathology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Prague, Czech Republic.

Department of Obstetrics and Gynecology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Prague, Czech Republic.

出版信息

Transl Res. 2023 Oct;260:61-68. doi: 10.1016/j.trsl.2023.05.004. Epub 2023 May 25.

Abstract

Testing of microsatellite instability is not only used as a triage for possible Lynch syndrome, but also to predict immunotherapy treatment response. The aim of this study was to assess the frequency of mismatch repair deficiency (MMR-D)/microsatellite instability (MSI) in 400 cases of non-endometrioid ovarian tumors (high-grade serous, low-grade serous, mucinous and clear cell), to compare different methodological approaches of testing, and to assess the optimal approach for next generation sequencing (NGS) MSI testing. For all tumors, we evaluated immunohistochemical (IHC) expression of MMR proteins and assessed microsatellite markers by PCR-based method. Except for high-grade serous carcinoma, we correlated the findings of IHC and PCR with NGS-based MSI testing. We compared the results with somatic and germline mutation in MMR genes. Among the whole cohort, seven MMR-D cases, all clear cell carcinomas (CCC), were found. On PCR analysis, 6 cases were MSI-high and one was MSS. In all cases, mutation of an MMR gene was found; in 2 cases, the mutation was germline (Lynch syndrome). An additional 5 cases with a mutation in MMR gene(s) with MSS status and without MMR-D were identified. We further utilized sequence capture NGS for MSI testing. Employing 53 microsatellite loci provided high sensitivity and specificity. Our study shows that MSI occurs in 7% of CCC while it is rare or absent in other nonendometrioid ovarian neoplasms. Lynch syndrome was present in 2% of patients with CCC. However, some cases with MSH6 mutation can evade all testing methods, including IHC, PCR, and NGS-MSI.

摘要

微卫星不稳定性检测不仅可作为林奇综合征(Lynch syndrome)的初步筛查,也可预测免疫治疗的反应。本研究旨在评估 400 例非子宫内膜样卵巢肿瘤(高级别浆液性癌、低级别浆液性癌、黏液性癌和透明细胞癌)中错配修复缺陷(MMR-D)/微卫星不稳定(MSI)的频率,比较不同的检测方法,并评估下一代测序(NGS)MSI 检测的最佳方法。我们对所有肿瘤进行了 MMR 蛋白免疫组化(IHC)表达评估,并采用基于 PCR 的方法检测微卫星标志物。除高级别浆液性癌外,我们还将 IHC 和 PCR 结果与 NGS 检测的 MSI 结果进行了相关性分析。我们将这些发现与 MMR 基因的体细胞和种系突变进行了比较。在整个队列中,我们发现了 7 例 MMR-D 病例,均为透明细胞癌(CCC)。PCR 分析显示,6 例为 MSI-H,1 例为 MSS。所有病例均发现 MMR 基因的突变;其中 2 例为种系突变(林奇综合征)。我们还发现了另外 5 例 MMR 基因发生突变、状态为 MSS 且不存在 MMR-D 的病例。我们进一步利用序列捕获 NGS 进行 MSI 检测。使用 53 个微卫星位点提供了高灵敏度和特异性。我们的研究表明,MSI 发生在 7%的 CCC 中,而在其他非子宫内膜样卵巢肿瘤中较为罕见或不存在。林奇综合征在 2%的 CCC 患者中存在。然而,一些 MSH6 突变的病例可能逃避所有的检测方法,包括 IHC、PCR 和 NGS-MSI。

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