Immunohistochemical (IHC) quantification of estrogen receptor-α (ER) is used for assessment of treatment regimen in breast cancer. Different ER IHC assays may produce diverging results, because of different antibody clones, protocols, and stainer platforms. Objective tissue-based techniques to assess sensitivity and specificity of IHC assays are therefore needed. We tested the usability of ER mRNA-in situ hybridization (mRNA-ISH) in comparison with assays based on clones SP1 and 6F11. We selected 56 archival specimens according to their reported ER IHC positivity, representing a wide spectrum from negative to strongly positive cases. The specimens were used to prepare 4 TMAs with 112 cores. Serial sections of each TMA were stained for ER and pan-cytokeratin (PCK) by IHC and ESR1 (ER gene) by mRNA-ISH. Digital image analysis (DIA) was used to determine ER IHC H-score. ESR1 mRNA-ISH was scored both manually and by DIA. DIA showed a nonlinear correlation between IHC and ESR1 mRNA-ISH with R2-values of 0.80 and 0.78 for the ER antibody clones SP1 and 6F11, respectively. Comparison of manual mRNA-ISH scoring categories and SP1 and 6F11 IHC H-scores showed a highly significant relationship (P<0.001). In conclusion, the study showed good correlation between mRNA-ISH and IHC, suggesting that mRNA-ISH can be a valuable tool in the assessment of the sensitivity and specificity of ER IHC assays.

中文翻译：

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乳腺癌中的雌激素受体-α定量

雌激素受体-α (ER) 的免疫组织化学 (IHC) 定量用于评估乳腺癌的治疗方案。由于不同的抗体克隆、实验方案和染色平台，不同的 ER IHC 检测可能会产生不同的结果。因此，需要客观的基于组织的技术来评估 IHC 检测的敏感性和特异性。与基于克隆 SP1 和 6F11 的分析相比，我们测试了 ER mRNA 原位杂交 (mRNA-ISH) 的可用性。我们根据他们报告的 ER IHC 阳性选择了 56 个档案标本，代表了从阴性到强阳性病例的广泛范围。这些样本用于制备 4 个具有 112 个芯的 TMA。通过 IHC 对每个 TMA 的连续切片进行 ER 和泛细胞角蛋白 (PCK) 染色，通过 mRNA-ISH 对 ESR1（ER 基因）进行染色。数字图像分析 (DIA) 用于确定 ER IHC H 评分。ESR1 mRNA-ISH 由手动和 DIA 评分。DIA 显示 IHC 和 ESR1 mRNA-ISH 之间存在非线性相关性，ER 抗体克隆 SP1 和 6F11 的 R2 值分别为 0.80 和 0.78。手动 mRNA-ISH 评分类别与 SP1 和 6F11 IHC H 评分的比较显示出高度显着的关系（P<0.001）。总之，该研究显示 mRNA-ISH 和 IHC 之间具有良好的相关性，这表明 mRNA-ISH 可以成为评估 ER IHC 检测的敏感性和特异性的有价值的工具。手动 mRNA-ISH 评分类别与 SP1 和 6F11 IHC H 评分的比较显示出高度显着的关系（P<0.001）。总之，该研究显示 mRNA-ISH 和 IHC 之间具有良好的相关性，这表明 mRNA-ISH 可以成为评估 ER IHC 检测的敏感性和特异性的有价值的工具。手动 mRNA-ISH 评分类别与 SP1 和 6F11 IHC H 评分的比较显示出高度显着的关系（P<0.001）。总之，该研究显示 mRNA-ISH 和 IHC 之间具有良好的相关性，这表明 mRNA-ISH 可以成为评估 ER IHC 检测的敏感性和特异性的有价值的工具。