Standardization of Estrogen Receptor Measurement in Breast Cancer Suggests False-Negative Results Are a Function of Threshold Intensity Rather Than Percentage of Positive Cells
Standardization of Estrogen Receptor Measurement in Breast Cancer Suggests False-Negative Results Are a Function of Threshold Intensity Rather Than Percentage of Positive Cells
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DOI：
10.1200/jco.2010.32.9706
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发表时间：
2011-08-01
影响因子：
45.3
通讯作者：
Rimm, David L.
中科院分区：
文献类型：
--
作者：
Welsh, Allison W.;Moeder, Christopher B.;Rimm, David L.
目的最近的错误分类（假阴性）事件提高了人们对免疫组织化学 (IHC) 在评估乳腺癌雌激素受体 (ER) 方面的局限性的认识。在这里，我们定义了一种标准化 ER 测量的新方法，然后检查百分比和强度阈值（免疫反应性）的变化，以评估测试不一致的来源。方法开发了一种测定法，通过使用对照组织微阵列 (TMA) 和一系列细胞系来量化 ER，其中通过定量免疫印迹与定量免疫荧光 (QIF) 自动定量分析 (AQUA) 方法并行分析 ER 免疫反应性。该测定用于评估耶鲁大学两个独立回顾性队列中的 ER 蛋白表达阈值，并与传统方法进行比较。结果两种分析方法显示，阳性细胞百分比从 10% 变化至 1% 并没有显着影响 ER 阳性患者的总数。两个耶鲁大学 TMA 队列的标准化 ER 检测显示，67.9% 和 82.5% 的患者高于 2 pg/μ g 免疫反应阈值。我们发现，与病理学家的阈值判断相比，9.1%和19.7%的患者为QIF阳性/IHC阴性，4.0%和0.4%的患者为QIF阴性/IHC阳性，总共有13.1%和20.1%的病例存在差异。对两个队列的生存评估表明，QIF 阳性/病理学家阴性的患者的结果与两种检测均呈阳性的患者的结果相似。结论通过对两个独立队列使用定量、标准化检测来评估强度阈值，表明与当前 IHC 方法的不一致在 10% 至 20% 范围内，其中结果不一致的患者的预后结果与结果一致的 ER 阳性患者相似。
PurposeRecent misclassification (false negative) incidents have raised awareness concerning limitations of immunohistochemistry (IHC) in assessment of estrogen receptor (ER) in breast cancer. Here we define a new method for standardization of ER measurement and then examine both change in percentage and threshold of intensity (immunoreactivity) to assess sources for test discordance.MethodsAn assay was developed to quantify ER by using a control tissue microarray (TMA) and a series of cell lines in which ER immunoreactivity was analyzed by quantitative immunoblotting in parallel with the automated quantitative analysis (AQUA) method of quantitative immunofluorescence (QIF). The assay was used to assess the ER protein expression threshold in two independent retrospective cohorts from Yale and was compared with traditional methods.ResultsTwo methods of analysis showed that change in percentage of positive cells from 10% to 1% did not significantly affect the overall number of ER-positive patients. The standardized assay for ER on two Yale TMA cohorts showed that 67.9% and 82.5% of the patients were above the 2-pg/mu g immunoreactivity threshold. We found 9.1% and 19.7% of the patients to be QIF-positive/ IHC-negative, and 4.0% and 0.4% to be QIF-negative/IHC-positive for a total of 13.1% and 20.1% discrepant cases when compared with pathologists' judgment of threshold. Assessment of survival for both cohorts showed that patients who were QIF-positive/ pathologist-negative had outcomes similar to those of patients who had positive results for both assays.ConclusionAssessment of intensity threshold by using a quantitative, standardized assay on two independent cohorts suggests discordance in the 10% to 20% range with current IHC methods, in which patients with discrepant results have prognostic outcomes similar to ER-positive patients with concordant results.