Detection of the BRAF V600E mutation in serous ovarian tumors: a comparative analysis of immunohistochemistry with a mutation-specific monoclonal antibody and allele-specific PCR
Detection of the BRAF V600E mutation in serous ovarian tumors: a comparative analysis of immunohistochemistry with a mutation-specific monoclonal antibody and allele-specific PCR
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DOI：
10.1016/j.humpath.2012.07.010
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发表时间：
2013-03-01
期刊：
影响因子：
3.3
通讯作者：
Fend, Falko
中科院分区：
文献类型：
--
作者：
Boesmueller, Hans;Fischer, Anna;Fend, Falko
丝裂原活化蛋白激酶途径的组分突变，主要是BRAF，在浆液性卵巢交界性肿瘤中很常见，而高级别浆液性卵巢癌很少显示此特征。随着对BRAF突变的癌症有活性的特异性激酶抑制剂的出现，快速和灵敏地检测BRAF V600 E（迄今为止该基因最常见的突变）具有很大的实际意义。目前，BRAF突变通过基于DNA的技术检测。最近，已经描述了适用于存档组织的BRAF V600E蛋白特异性单克隆抗体（VE 1）。在这项研究中，我们比较了V600E突变检测浆液性卵巢肿瘤的VE1免疫染色和等位基因特异性聚合酶链反应。所有141例高级别浆液性卵巢癌均显示阴性或罕见弱的弥漫性背景VE 1免疫染色，所有检测病例均通过分子分析证实BRAF野生型。相比之下，7例低级别浆液性癌中1例（14%）和31例浆液性交界性肿瘤中22例（71%）显示中度至强阳性。在所有肿瘤细胞充足的病例中，免疫染色均清晰可评价，只有极少数细胞质狭窄的病例难以解释。V600 E突变证实等位基因特异性聚合酶链反应和测序在所有VE 1阳性病例。两例VE1阳性病例的上皮细胞含量低，需要重复显微解剖，以确认突变的存在。VE1抗体的免疫组化是检测浆液性卵巢肿瘤中BRAF V600E突变的特异性和敏感性工具，并可提供实用的筛选试验，特别是在上皮含量低的肿瘤样本中。(c)2013 Elsevier Inc. All rights reserved.
Mutations of components of the mitogen-activated protein kinase pathway, mainly BRAF, are common in serous ovarian borderline tumors, whereas high-grade serous ovarian carcinomas rarely show this feature. With the advent of specific kinase inhibitors active against BRAF-mutated cancers, rapid and sensitive detection of the BRAF V600E, by far the most common mutation of this gene, is of great practical relevance. Currently, BRAF mutations are detected by DNA-based techniques. Recently, a monoclonal antibody (VE1) specific for the BRAF V600E protein suitable for archival tissues has been described. In this study, we compared detection of the V600E mutation in serous ovarian tumors by VE1 immunostaining and by allele-specific polymerase chain reaction. All 141 cases of high-grade serous ovarian cancer showed negative or rarely weak, diffuse background VE1 immunostaining, and BRAF wild type was confirmed by molecular analysis in all tested cases. In contrast, 1 (14%) of 7 low-grade serous carcinomas and 22 (71%) of 31 serous borderline tumors revealed moderate to strong VE1 positivity. Immunostaining was clearly evaluable in all cases with sufficient tumor cells, and only rare cases with narrow cytoplasm were difficult to interpret. The V600E mutation was confirmed by allele-specific polymerase chain reaction and sequencing in all VE1-positive cases. Two VE1-positive cases with low epithelial cell content required repeat microdissection to confirm the presence of the mutation. Immunohistochemistry with the VE1 antibody is a specific and sensitive tool for detection of the BRAF V600E mutation in serous ovarian tumors and may provide a practical screening test, especially in tumor samples with low epithelial content. (c) 2013 Elsevier Inc. All rights reserved.