Naber S P, Tsutsumi Y, Yin S, Zolnay S A, Mobtaker H, Marks P J, McKenzie S J, DeLellis R A, Wolfe H J
Department of Pathology, Tufts University, New England Medical Center Hospital, Boston, Massachusetts 02111.
Am J Clin Pathol. 1990 Aug;94(2):125-36. doi: 10.1093/ajcp/94.2.125.
The development of a consistent strategy for the analysis of oncogene expression at the cellular level is essential for understanding the roles of these genes in the development and progression of human neoplasia. Detection of the neu oncogene products in breast carcinoma was selected as a model for analysis of oncogene expression. Fifty-two primary human breast carcinomas were evaluated by quantitation of neu DNA amplification and mRNA expression and by localization of neu mRNA and protein (p 185) at the cellular level by in situ hybridization (ISH) and immunohistochemistry (IHC). The specificity and sensitivity of the molecular and immunologic probes for neu were established with the use of genetically engineered cell lines that overexpressed either neu or epidermal growth factor receptor (EGFR). Twenty-nine percent of breast carcinomas demonstrated neu DNA amplification and mRNA overexpression, and there was close correlation between the level of neu mRNA expression and detection of neu gene products by ISH and IHC. Thirty-two percent of carcinomas demonstrated neu mRNA overexpression by ISH. The immunohistochemical method using TA1 monoclonal antibody for p185 was exquisitely sensitive in acetone-fixed frozen sections and provided an excellent approach for judging overexpression as confirmed by the various molecular analyses. All areas of nonmalignant breast epithelium stained weakly, and a wide range of staining intensity was observed in malignant breast epithelium, with 31% of carcinomas judged to be p185 overexpressors. Heterogeneous expression of p185 was seen in some carcinomas. This study provides a strategic approach for the evaluation of oncogene expression in human tumors.
制定一种在细胞水平分析癌基因表达的连贯策略对于理解这些基因在人类肿瘤发生和发展中的作用至关重要。检测乳腺癌中的neu癌基因产物被选作分析癌基因表达的模型。通过定量neu DNA扩增和mRNA表达，以及通过原位杂交（ISH）和免疫组织化学（IHC）在细胞水平定位neu mRNA和蛋白（p185），对52例原发性人类乳腺癌进行了评估。使用过表达neu或表皮生长因子受体（EGFR）的基因工程细胞系确定了neu分子和免疫探针的特异性和敏感性。29%的乳腺癌显示neu DNA扩增和mRNA过表达，并且neu mRNA表达水平与ISH和IHC检测neu基因产物之间存在密切相关性。32%的癌通过ISH显示neu mRNA过表达。使用针对p185的TA1单克隆抗体的免疫组织化学方法在丙酮固定的冰冻切片中极其敏感，并为判断过表达提供了一种极好的方法，各种分子分析均证实了这一点。所有非恶性乳腺上皮区域染色较弱，在恶性乳腺上皮中观察到广泛的染色强度范围，31%的癌被判定为p185过表达者。在一些癌中可见p185的异质性表达。本研究为评估人类肿瘤中的癌基因表达提供了一种策略性方法。
