McManus D T, Patterson A H, Maxwell P, Humphreys M W, Anderson N H
Immunohistochemistry and Molecular Pathology Laboratory, Department of Pathology, Belfast, Northern Ireland, UK.
Mol Pathol. 1999 Apr;52(2):75-7. doi: 10.1136/mp.52.2.75.
To develop a method for the detection of amplification of the erbB2 oncogene in breast cancer fine needle aspirates using fluorescence in situ hybridisation (FISH) and to compare amplification with immunohistochemical detection of the erbB2 protein.
A digoxigenin labelled probe to the erbB2 gene was hybridised to 15 aspirates prepared from operative breast cancer specimens. A chromosome 17 centromere probe was also hybridised to the aspirates either separately or in combination with the erbB2 probe. The aspirates were scored for erbB2 amplification and chromosome 17 centromere number. Subsequently, paraffin wax embedded sections of the tumours were stained with the antibody CB11 and scored for the presence of membrane staining.
Three of the 15 tumour aspirates showed high level amplification of erbB2 detected by FISH. These three tumours also showed chromosome 17 polysomy and diffuse membrane staining by immunohistochemistry.
FISH can be used to detect erbB2 amplification in fine needle aspirates and results correlate with conventional immunohistochemical staining. Difficulties were encountered in the visualisation of the signals in non-amplified cases without the use of specialised digital imaging.
开发一种使用荧光原位杂交(FISH)检测乳腺癌细针穿刺抽吸物中erbB2癌基因扩增的方法,并将扩增情况与erbB2蛋白的免疫组织化学检测结果进行比较。
将地高辛标记的erbB2基因探针与从手术切除的乳腺癌标本制备的15份抽吸物进行杂交。还将17号染色体着丝粒探针单独或与erbB2探针联合与抽吸物进行杂交。对抽吸物进行erbB2扩增和17号染色体着丝粒数量评分。随后,用抗体CB11对肿瘤的石蜡包埋切片进行染色,并对膜染色情况进行评分。
15份肿瘤抽吸物中有3份通过FISH检测显示erbB2高水平扩增。这3个肿瘤还显示17号染色体多体性,并且免疫组织化学显示弥漫性膜染色。
FISH可用于检测细针穿刺抽吸物中的erbB2扩增,结果与传统免疫组织化学染色相关。在不使用专门数字成像的情况下,非扩增病例的信号可视化存在困难。