Evaluation of epidermal growth factor receptor (EGFR) by chromogenic in situ hybridization (CISH™) and immunohistochemistry (IHC) in archival gliomas using bright-field microscopy
Evaluation of epidermal growth factor receptor (EGFR) by chromogenic in situ hybridization (CISH™) and immunohistochemistry (IHC) in archival gliomas using bright-field microscopy
复制标题
DOI：
10.1097/00019606-200403000-00001
复制
发表时间：
2004-03-01
影响因子：
--
通讯作者：
Shi, ZR
中科院分区：
文献类型：
--
作者：
Marquez, A;Wu, R;Shi, ZR
继发于EGFR基因扩增的EGFR过表达是原发性恶性胶质瘤的常见特征。为了正确评估EGFR蛋白和基因水平作为神经胶质瘤中可能的预后和预测标志物，可以在病理学实验室中常规使用以评估EGFR状态的直接测定变得至关重要。应用地高辛标记的埃格和生物素标记的7号染色体着丝粒探针，通过发色原位杂交（CISH（TM））检测了34例福尔马林固定、石蜡包埋的良性和恶性胶质瘤中EGFR基因扩增和7号染色体非整倍体。在40倍物镜透镜下通过明视野显微镜评价结果。采用单克隆抗体31G7免疫组化检测EGFR蛋白水平。5例，3例星形细胞瘤III级（33%）和2例多形性胶质母细胞瘤（GBM）（33%），EGFR扩增显示为二氨基联苯胺染色的多个点，提示双分钟染色体的模式。7号染色体多体性见于68%的胶质瘤、100%的胶质母细胞瘤、67%的星形细胞瘤III级、42%的星形细胞瘤II级、50%的星形细胞瘤1级、100%的室管膜瘤和1例混合性胶质瘤III级。62%的胶质瘤中存在EGFR蛋白的高表达，并表现为细胞膜和细胞质染色。所有EGFR基因扩增的肿瘤均呈EGFR高表达。在所有级别的胶质瘤中均观察到埃格的高表达而无基因扩增。同时检测EGFR基因拷贝或7号染色体着丝粒信号沿着组织形态学使我们能够比较CISH（TM）结果与IHC结果。我们的研究结果表明，CISH（TM）是一种客观、实用、准确的检测胶质瘤EGFR基因状态的方法。
Overexpression of EGFR secondary to EGFR gene amplification is a common feature in primary malignant gliomas. To correctly assess EGFR protein and gene level as possible prognostic and predictive markers in gliomas, straightforward assays, which can be used routinely in the pathology laboratory to evaluate EGFR status, becomes critical. EGFR gene amplification and chromosome 7 aneuploidy was detected in 34 formalin-fixed, paraffin-embedded benign and malignant gliomas by chromogenic in situ hybridization (CISH(TM)) using digoxigenin-labeled EGER and biotin-labeled chromosome 7 centromeric probes. The results were evaluated by bright-field microscopy under a 40x objective lens. EGFR protein level was detected by immunohistochemistry (IHC) using monoclonal antibody 31G7. Five cases, 3 astrocytoma grade III (33%) and 2 glioblastoma multiforme (GBM) (33%), had EGFR amplification displayed as diaminobenzidine-stained multiple dots suggesting the pattern of double-minute chromosomes. Chromosome 7 polysomy was found in 68% gliomas, 100% GBM, 67% astrocytoma grade III, 42% astrocytoma grade II, 50% astrocytoma grade 1, 100% ependymoma, and the I case of mixed glioma III. High expression of EGFR protein was present in 62% gliomas and displayed membrane and cytoplasmic staining. All tumors with EGFR gene amplification showed EGFR high expression. High expression of EGER without gene amplification was observed in all grades of gliomas. Simultaneous detection of EGFR gene copies or chromosome 7 centromere signals along with tissue morphology allows us to compare CISH(TM) results easily with IHC results. Our results show that CISH(TM) is an objective, practical, and accurate assay to screen for EGFR gene status in gliomas.