探索IHC和下一代测序方法在子宫内膜癌检测和分类中的诊断潜力

Azhin Saber Ali, Lana Sardar Alalem
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摘要

背景:在全球范围内,2018年子宫内膜癌(EC)被认为是第六大最常诊断的癌症,也是第14大癌症原因,女性死亡人数更多。如今,经典和现代分类可以用于诊断各种类型的癌症,包括子宫内膜癌。基于此,我们的研究目的是确定免疫组织化学观察的有效性,这是一种基于现代分类的快速简便的确定MSI和/或其他类型EC的方法。由于分子研究的成本较高,我们想比较两种方法,并展示用免疫组化代替分子研究的可能性。方法:本研究旨在建立针对12个基因的子宫内膜癌患者Next Generation Sequencing NGS小基因面板;MSH6、MLH3、MSH3、CDH1、PTEN、MSH2、MLH1、PMS2、FGFR2、BRCA1、MUTYH和are1。采用DNBSEQ-G400平台,使用Human Core Exome试剂盒和Python软件进行分析。同时,Dako试剂盒对MSH6、MSH2、MLH、PMS2、PTEN、CDH1等6种一抗进行免疫组化检测。一抗应用于5µm福尔马林固定石蜡包埋(FFPE)。结果;组织病理学检查显示所有患者均为II期子宫内膜样癌。FIGO分级为Ia和Ib。免疫组化观察微卫星不稳定性(Microsatellite instability, MSI)、PTEN抗体核染色免疫反应和膜性E- cadherin免疫反应。分子研究发现了一些具有临床意义的多态性,其中一些有相互冲突的解释。结论;我们认为,简单的免疫反应染色程序可以作为MSI表型检测的替代方法,而不是任何类型的更昂贵和复杂的NGS方法。对于PTEN和ECAD1抗体的反应,在无法进行分子诊断的情况下,也可以进行可靠的诊断和治疗。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Exploring the Diagnostic Potential of IHC and Next-Generation Sequencing Approaches for Endometrial Carcinoma Detection and Classification
Background: Globally, in 2018 endometrial cancer (EC) regarded as the sixth most commonly diagnosed cancer and the fourteenth leading cause of cancer with more deaths in females. Nowadays classical and modern classifications can be performing for diagnosing various types of cancers including endometrial cancer. Based on this our Objective of study is to determine the effectiveness of immunohistochemical observation, a quick and easy method for determining MSI and/or other types of EC which based on modern classification. As molecular study takes more cost we want to compare the both methods and showing the possibility of doing IHC instead of molecular study. Methods: This study designed to establish a small gene panel of Next Generation Sequencing NGS of endometrial cancer patients targeting 12 genes; MSH6, MLH3, MSH3, CDH1, PTEN, MSH2, MLH1, PMS2, FGFR2, BRCA1, MUTYH and AREL1. By using DNBSEQ-G400 Platform, Human Core Exome kit and Python software for analysis is used. At the same time, Dako kit used to performing IHC for six primary antibodies used to detect each of MSH6, MSH2, MLH, PMS2, PTEN and CDH1. The primary antibodies were applied on 5 µm formalin- fixed paraffin- embedded (FFPE). Results; histopathological examinations showed that all patients were at stage II endometrioid endometrial carcinoma. The FIGO classification were Ia and Ib. Microsatellite instability (MSI), nuclear staining immunoreaction for PTEN antibodies and membranous E- cadherin immunoreactions observed through IHC study. The molecular studies detected several polymorphisms which have clinical significant and some of them have conflict interpretations. Conclusions; we considered that, simply immunoreaction staining procedure can be use as alternative method for MSI phenotype detection rather than any type of more expensive and complex method of NGS. Concerning PTEN and ECAD1 antibodies reactions, also could be dependable in diagnosis and treatment and performing them if molecular diagnosis not available.
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