Smith K L, Robbins P D, Dawkins H J, Papadimitriou J M, Redmond S L, Carrello S, Harvey J M, Sterrett G F
Department of Pathology, University of Western Australia, Nedlands.
Hum Pathol. 1994 Apr;25(4):413-8. doi: 10.1016/0046-8177(94)90152-x.
We describe a sensitive and practical in situ hybridization method, using a digoxigenin-labeled probe, for the detection of c-erbB-2 amplification in breast cancer in formalin-fixed, paraffin-embedded tissue sections. Forty-six primary breast carcinomas were studied. Nuclear hybridization signal was observed in 36 of 46 carcinomas. Signal was confined to malignant cells. Normal breast epithelium and stromal and inflammatory cells were uniformally negative. DNase predigestion, no-probe preparations, and competitive hybridization confirmed the specificity of the reaction. The hybridization reaction was localized to multiple discrete foci in tumor cell nuclei, suggesting multiple sites of gene copy and transcriptional activity in the nucleus. Considerable cell-to-cell variation in hybridization signal was evident within individual tumors and positive reactions were observed in several cases in which amplification could not be detected by either Southern or slot blot analysis. The high sensitivity and specificity of the reaction and its use in a tissue-based system will allow the study of a range of possible precursor lesions of breast cancer for evidence of c-erbB-2 amplification.
我们描述了一种灵敏且实用的原位杂交方法，该方法使用地高辛标记的探针，用于检测福尔马林固定、石蜡包埋的组织切片中乳腺癌组织的c-erbB-2基因扩增情况。我们研究了46例原发性乳腺癌。在46例癌组织中，有36例观察到核杂交信号。信号局限于恶性细胞。正常乳腺上皮细胞、基质细胞及炎性细胞均呈阴性。DNA酶预消化、无探针制剂及竞争性杂交证实了反应的特异性。杂交反应定位于肿瘤细胞核内多个离散的位点，提示细胞核内基因拷贝和转录活性的多个位点。在单个肿瘤内，杂交信号在细胞间存在明显差异，并且在几例通过Southern印迹或狭缝印迹分析均未检测到扩增的病例中观察到了阳性反应。该反应的高灵敏度和特异性及其在组织系统中的应用，将有助于研究一系列可能的乳腺癌前病变中c-erbB-2基因扩增的证据。
