Juarez Ignacio, Toro-Fernandez Juan Francisco, Vaquero-Yuste Christian, Molina-Alejandre Marta, Lasa Inmaculada, Gomez Remedios, Lopez Adela, Martin-Villa Jose Manuel, Gutierrez Alberto
Department of Immunology, Ophthalmology and ENT, Facultad de Medicina, Universidad Complutense de Madrid (UCM), 28040 Madrid, Spain.
Instituto de Investigación Sanitaria Gregorio Marañón (IiSGM), 28007 Madrid, Spain.
Biology (Basel). 2021 Jun 10;10(6):516. doi: 10.3390/biology10060516.
We have applied two PCR techniques, differential PCR (diffPCR) and qPCR for the identification of HER2 gene amplifications in genomic DNA of tumor and distal gastric samples from patients with gastric cancer. The diffPCR technique consists of the simultaneous amplification of the HER2 gene and a housekeeping gene by conventional PCR and the densitometric analysis of the bands obtained. We established a cut-off point based on the mean and standard deviation analyzing the DNA of 30 gastric tissues from patients undergoing non-cancer gastrectomy. diffPCR and qPCR yielded consistent results. HER2-overexpression was detected in 25% of patients and was further confirmed by immunohistochemistry and immunofluorescence. The approaches herein described may serve as complementary and reliable methods to assess HER2 amplification.
我们应用了两种聚合酶链式反应（PCR）技术，即差异PCR（diffPCR）和定量PCR（qPCR），来鉴定胃癌患者肿瘤和远端胃样本基因组DNA中的人表皮生长因子受体2（HER2）基因扩增情况。差异PCR技术包括通过常规PCR同时扩增HER2基因和一个管家基因，并对所得条带进行光密度分析。我们通过分析30例接受非癌性胃切除术患者的胃组织DNA，基于均值和标准差确定了一个临界值。差异PCR和定量PCR得出了一致的结果。在25%的患者中检测到HER2过表达，并通过免疫组织化学和免疫荧光进一步证实。本文所述方法可作为评估HER2扩增的补充且可靠的方法。
