非小细胞肺癌中表皮生长因子受体的蛋白过表达和基因扩增：四种市售抗体的免疫组织化学和荧光原位杂交研究比较。

Protein overexpression and gene amplification of epidermal growth factor receptor in nonsmall cell lung carcinomas: Comparison of four commercially available antibodies by immunohistochemistry and fluorescence in situ hybridization study.

机构信息

Department of Pathology, Seoul National University College of Medicine, 28 Yongon-dong, Chongno-gu, Seoul 110-799, Republic of Korea.

出版信息

Lung Cancer. 2010 Jun;68(3):375-82. doi: 10.1016/j.lungcan.2009.07.014. Epub 2009 Aug 26.

DOI:10.1016/j.lungcan.2009.07.014

PMID:19712993

Abstract

Epidermal growth factor receptor (EGFR) overexpression in nonsmall cell lung carcinomas (NSCLC) is variable ranging from 19% to 89% and its prognostic value remains controversial. We tried to investigate (1) EGFR protein expression using four different antibodies, (2) the correlation between protein overexpression and EGFR gene amplification, and (3) the correlation between EGFR genetic status and clinicopathologic features in NSCLC. We examined EGFR protein expression using four different antibodies including Zymed EGFR kit (Clone 31G7), Dako EGFR pharmDx kit (Clone 2-18C9), Dako (Clone H11) and Novocastra (Clone EGFR 113) by immunohistochemical analysis. The protein overexpression was compared to gene amplification status by fluorescence in situ hybridization (FISH). EGFR protein overexpression was observed in 56% of tumors with Zymed EGFR kit, in 51% with Dako EGFR pharmDx kit, in 5% with Dako and in 18% with Novocastra (p=0.010). Both Zymed and Dako pharmDx kit were more sensitive than the Dako test (clone H11) and Novocastra clone EGFR 113. EGFR overexpression was more prominent in squamous cell carcinomas (SCC) than adenocarcinomas (ADC) (71% vs. 48%, p=0.001 with Zymed, 61% vs. 45%, p=0.011 with Dako pharmDx kit; respectively). EGFR FISH-positivity as represented by high polysomy and gene amplification was observed in 45% of the NSCLC patients. Protein expression levels significantly correlated with the gene copy number per tumor cell (p<0.001). Our data showed a higher percentage of positive cells detected by Zymed and Dako pharmDx tests. The EGFR protein overexpression rate varied from 4% to 72% according to different antibody clones and histologic types. EGFR protein expression detected by Zymed and Dako pharmDx was significantly associated with a high EGFR gene copy number.

摘要

表皮生长因子受体 (EGFR) 在非小细胞肺癌 (NSCLC) 中的过度表达率为 19%至 89%，其预后价值仍存在争议。我们试图研究 (1) 使用四种不同抗体的 EGFR 蛋白表达，(2) 蛋白过表达与 EGFR 基因扩增之间的相关性，以及 (3) NSCLC 中 EGFR 遗传状态与临床病理特征之间的相关性。我们使用四种不同的抗体，包括 Zymed EGFR 试剂盒 (克隆 31G7)、Dako EGFR pharmDx 试剂盒 (克隆 2-18C9)、Dako (克隆 H11) 和 Novocastra (克隆 EGFR 113) 通过免疫组织化学分析来检测 EGFR 蛋白表达。蛋白过表达与荧光原位杂交 (FISH) 检测的基因扩增状态进行比较。使用 Zymed EGFR 试剂盒检测到 56%的肿瘤存在 EGFR 蛋白过表达，使用 Dako EGFR pharmDx 试剂盒检测到 51%的肿瘤存在 EGFR 蛋白过表达，使用 Dako 试剂盒检测到 5%的肿瘤存在 EGFR 蛋白过表达，使用 Novocastra 试剂盒检测到 18%的肿瘤存在 EGFR 蛋白过表达 (p=0.010)。Zymed 和 Dako pharmDx 试剂盒均比 Dako 试剂盒 (克隆 H11) 和 Novocastra 克隆 EGFR 113 更敏感。EGFR 过表达在鳞状细胞癌 (SCC) 中比在腺癌 (ADC) 中更为明显 (Zymed：71% vs. 48%，p=0.001；Dako pharmDx 试剂盒：61% vs. 45%，p=0.011)。EGFR FISH 阳性(以高倍体和基因扩增表示)在 45%的 NSCLC 患者中观察到。蛋白表达水平与每个肿瘤细胞的基因拷贝数显著相关 (p<0.001)。我们的数据显示，Zymed 和 Dako pharmDx 检测到的阳性细胞百分比更高。根据不同的抗体克隆和组织学类型，EGFR 蛋白过表达率从 4%到 72%不等。Zymed 和 Dako pharmDx 检测到的 EGFR 蛋白表达与高 EGFR 基因拷贝数显著相关。