Hunter S B, Abbott K, Varma V A, Olson J J, Barnett D W, James C D
Department of Pathology, Emory University School of Medicine, Atlanta, Georgia 30322.
J Neuropathol Exp Neurol. 1995 Jan;54(1):57-64. doi: 10.1097/00005072-199501000-00007.
A series of 43 human gliomas, consisting of 30 glioblastomas, 7 anaplastic astrocytomas, 3 low grade astrocytomas, 2 ependymomas, and 1 oligodendroglioma, was studied for amplification of the epidermal growth factor receptor (EGFR) and mouse double minute 2 (MDM2) genes. DNA extracted from formalin-fixed, paraffin-embedded tissue sections was analyzed by differential PCR and the results were compared with slot blot examination of DNA extracted from frozen tissue from the same neoplasms. Twelve glioblastomas (40%) showed amplification of the EGFR gene, and overexpression of EGFR was evident in each of these tumors as indicated by the immunoperoxidase technique. Two of the tumors with EGFR gene amplification also revealed amplification of the MDM2 gene, while one additional glioblastoma revealed MDM2 amplification only. A 100% concordance in the detection of amplification was observed between differential PCR and slot blot analysis; consequently, these results indicate that differential PCR using DNA extracted from archival tissue sections is a reliable method of demonstrating gene amplifications in glial tumors.
对一系列43例人类胶质瘤进行了研究，其中包括30例胶质母细胞瘤、7例间变性星形细胞瘤、3例低级别星形细胞瘤、2例室管膜瘤和1例少突胶质细胞瘤，以检测表皮生长因子受体（EGFR）和小鼠双微体2（MDM2）基因的扩增情况。从福尔马林固定、石蜡包埋的组织切片中提取DNA，通过差异PCR进行分析，并将结果与从同一肿瘤的冷冻组织中提取的DNA进行斑点杂交检测结果相比较。12例胶质母细胞瘤（40%）显示EGFR基因扩增，免疫过氧化物酶技术表明，这些肿瘤中的每一例均有明显的EGFR过表达。2例EGFR基因扩增的肿瘤也显示MDM2基因扩增，而另有1例胶质母细胞瘤仅显示MDM2扩增。差异PCR和斑点杂交分析在扩增检测方面的一致性为100%；因此，这些结果表明，使用从存档组织切片中提取的DNA进行差异PCR是一种可靠的方法，可用于证明胶质肿瘤中的基因扩增。
