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鉴定与 Epstein-Barr 病毒相关的胃癌中特异性甲基化的基因。

Identification of genes specifically methylated in Epstein-Barr virus-associated gastric carcinomas.

机构信息

Department of Oncology and Laboratory Medicine, Yamaguchi University Graduate School of Medicine, Ube, Japan.

出版信息

Cancer Sci. 2013 Oct;104(10):1309-14. doi: 10.1111/cas.12228. Epub 2013 Aug 9.

Abstract

We studied the comprehensive DNA methylation status in the naturally derived gastric adenocarcinoma cell line SNU-719, which was infected with the Epstein-Barr virus (EBV) by methylated CpG island recovery on chip assay. To identify genes specifically methylated in EBV-associated gastric carcinomas (EBVaGC), we focused on seven genes, TP73, BLU, FSD1, BCL7A, MARK1, SCRN1, and NKX3.1, based on the results of methylated CpG island recovery on chip assay. We confirmed DNA methylation of the genes by methylation-specific PCR and bisulfite sequencing in SNU-719. The expression of the genes, except for BCL7A, was upregulated by a combination of 5-Aza-2'-deoxycytidine and trichostatin A treatment in SNU-719. After the treatment, unmethylated DNA became detectable in all seven genes by methylation-specific PCR. We verified DNA methylation of the genes in 75 primary gastric cancer tissues from 25 patients with EBVaGC and 50 EBV-negative patients who were controls. The methylation frequencies of TP73, BLU, FSD1, BCL7A, MARK1, SCRN1, and NKX3.1 were significantly higher in EBVaGC than in EBV-negative gastric carcinoma. We identified seven genes with promoter regions that were specifically methylated in EBVaGC. Inactivation of these genes may suppress their function as tumor suppressor genes or tumor-associated antigens and help to develop and maintain EBVaGC.

摘要

我们研究了自然衍生的胃腺癌细胞系 SNU-719 的综合 DNA 甲基化状态,该细胞系通过芯片分析中的甲基化 CpG 岛恢复感染了 Epstein-Barr 病毒 (EBV)。为了鉴定与 EBV 相关的胃癌 (EBVaGC) 中特异性甲基化的基因,我们根据甲基化 CpG 岛恢复芯片分析的结果,重点关注了七个基因,包括 TP73、BLU、FSD1、BCL7A、MARK1、SCRN1 和 NKX3.1。我们通过甲基化特异性 PCR 和 SNU-719 中的亚硫酸氢盐测序确认了这些基因的 DNA 甲基化。除了 BCL7A 之外,这些基因的表达在 SNU-719 中通过 5-Aza-2'-脱氧胞苷和曲古抑菌素 A 的联合处理而上调。处理后,通过甲基化特异性 PCR 可以检测到所有七个基因的非甲基化 DNA。我们在 25 例 EBVaGC 患者和 50 例 EBV 阴性对照患者的 75 例原发性胃癌组织中验证了这些基因的 DNA 甲基化。TP73、BLU、FSD1、BCL7A、MARK1、SCRN1 和 NKX3.1 的甲基化频率在 EBVaGC 中明显高于 EBV 阴性胃癌。我们鉴定了七个在 EBVaGC 中启动子区域特异性甲基化的基因。这些基因的失活可能会抑制其作为肿瘤抑制基因或肿瘤相关抗原的功能,并有助于 EBVaGC 的发生和维持。

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