Arbogast Stefanie, Behnke Silvia, Opitz Isabelle, Stahel Rolf A, Seifert Burkhardt, Weder Walter, Moch Holger, Soltermann Alex
Institute for Surgical Pathology, Department of Pathology, University Hospital Zurich, Zurich, Switzerland.
Appl Immunohistochem Mol Morphol. 2011 Mar;19(2):99-105. doi: 10.1097/PAI.0b013e3181f1feeb.
The excision repair cross-complementation group 1 (ERCC1) protein is the key enzyme of the nucleotide excision repair (NER) pathway and thus a potential predictor for platinum-based chemotherapy response. We aimed for evaluating different anti-ERCC1 antibodies on formalin-fixed tumor tissue of non-small cell lung cancer patients by automated immunohistochemistry (IHC).
ERCC1 protein expression was assessed on a tissue microarray of 491 NSCLC's using 2 monoclonal mouse (Mab 8F1, Mab D-10) and 1 polyclonal rabbit (Rab FL-297) antibody. Two automated IHC platforms with different detection systems and immunofluorescence were used. Protein expression levels were independently scored by 2 pathologists for both intensity and intensity multiplied by percentage of positive cells (H-score).
On both platforms, the 8F1 ab showed best nuclear staining quality. D-10 had additional unspecific background at the plasma membrane and in goblet cells. FL-297 could not be scored owing to high cytoplasmic background. Both 8F1 and D-10 antibodies produced a speckled granular pattern over the whole nuclear compartment. No intranuclear compartmentalization was observed, apart from omission of the nucleolus. Interobserver κ value was good to very good for 8F1 and D-10. Using 8F1, low ERCC1 was correlated with the adenocarcinoma histotype, increased tumor size and clinical stage, high pT and pN category and the presence of metastasis. No relation to progression-free or overall survival was observed.
In terms of staining quality and restriction to the nuclear compartment, the antibody 8F1 is superior to D-10 or FL-297 on automated IHC platforms.
切除修复交叉互补基因1（ERCC1）蛋白是核苷酸切除修复（NER）途径的关键酶，因此是基于铂类化疗反应的潜在预测指标。我们旨在通过自动免疫组织化学（IHC）评估不同抗ERCC1抗体对非小细胞肺癌患者福尔马林固定肿瘤组织的作用。
使用2种单克隆小鼠抗体（Mab 8F1、Mab D-10）和1种多克隆兔抗体（Rab FL-297）在491例非小细胞肺癌组织微阵列上评估ERCC1蛋白表达。使用了两个具有不同检测系统和免疫荧光的自动IHC平台。由2名病理学家独立对蛋白表达水平的强度以及强度乘以阳性细胞百分比（H评分）进行评分。
在两个平台上，8F1抗体均显示出最佳的细胞核染色质量。D-10在质膜和杯状细胞中有额外的非特异性背景。由于高细胞质背景，FL-297无法进行评分。8F1和D-10抗体在整个核区均产生斑点状颗粒模式。除了核仁缺失外，未观察到核内分区。观察者间对8F1和D-10的κ值为良好至非常好。使用8F1时，低ERCC1与腺癌组织学类型、肿瘤大小增加和临床分期、高pT和pN类别以及转移的存在相关。未观察到与无进展生存期或总生存期的关系。
在染色质量和对核区的限制方面，8F1抗体在自动IHC平台上优于D-10或FL-297。
