The reliability of immunohistochemistry as a prescreening method for the diagnosis of hereditary nonpolyposis colorectal cancer (HNPCC)--results of an international collaborative study.
The reliability of immunohistochemistry as a prescreening method for the diagnosis of hereditary nonpolyposis colorectal cancer (HNPCC)--results of an international collaborative study.
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DOI：
10.1023/a:1013840907881
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发表时间：
2001-01-01
期刊：
影响因子：
2.2
通讯作者：
Moslein, G
中科院分区：
文献类型：
--
作者：
Muller, W;Burgart, L J;Moslein, G
遗传性非息肉病性结直肠癌综合征 (HNPCC) 是一种常染色体显性遗传疾病，占所有结直肠癌的 2-5%，以及一小部分子宫内膜癌、上尿路癌和其他胃肠道癌。检测 HNPCC 潜在缺陷（DNA 错配修复酶失活）的检测将有助于识别 HNPCC 先证者。针对 hMLH1 和 hMSH2（大多数 HNPCC 癌症的两种 DNA 错配修复蛋白）的单克隆抗体已在市场上出售。本研究旨在研究这些抗体在确定石蜡包埋福尔马林固定组织中这些蛋白质的表达状态方面的潜在效用，并确定与成功染色相关的关键技术方案组成部分。一组 20 例已知 hMLH1 和 hMSH2 突变和表达状态的结直肠癌病例在多个机构进行了免疫过氧化物酶染色，每个机构都使用自己的技术方案。 hMSH2 染色在大多数实验室中都是成功的，而 hMLH1 染色在多个实验室中被证明存在问题。然而，少数实验室表现出了优异的结果，包括对两种单克隆抗体的高辨别力。这些实验室以高灵敏度和特异性正确鉴定了 hMLH1 或 hMSH2 失活。与成功染色相关的关键方案点是抗原修复步骤，涉及热处理和 EDTA 或柠檬酸盐缓冲液。这项研究证明了免疫组织化学在检测 HNPCC 先证者中的潜在用途，并确定了成功染色的关键技术组件。
Hereditary nonpolyposis colorectal cancer syndrome (HNPCC) is an autosomal dominant condition accounting for 2-5% of all colorectal carcinomas as well as a small subset of endometrial, upper urinary tract and other gastrointestinal cancers. An assay to detect the underlying defect in HNPCC, inactivation of a DNA mismatch repair enzyme, would be useful in identifying HNPCC probands. Monoclonal antibodies against hMLH1 and hMSH2, two DNA mismatch repair proteins which account for most HNPCC cancers, are commercially available. This study sought to investigate the potential utility of these antibodies in determining the expression status of these proteins in paraffin-embedded formalin-fixed tissue and to identify key technical protocol components associated with successful staining. A set of 20 colorectal carcinoma cases of known hMLH1 and hMSH2 mutation and expression status underwent immunoperoxidase staining at multiple institutions, each of which used their own technical protocol. Staining for hMSH2 was successful in most laboratories while staining for hMLH1 proved problematic in multiple labs. However, a significant minority of laboratories demonstrated excellent results including high discriminatory power with both monoclonal antibodies. These laboratories appropriately identified hMLH1 or hMSH2 inactivation with high sensitivity and specificity. The key protocol point associated with successful staining was an antigen retrieval step involving heat treatment and either EDTA or citrate buffer. This study demonstrates the potential utility of immunohistochemistry in detecting HNPCC probands and identifies key technical components for successful staining.