The effect of EBV on WIF1, NLK, and APC gene methylation and expression in gastric carcinoma and nasopharyngeal cancer
The effect of EBV on WIF1, NLK, and APC gene methylation and expression in gastric carcinoma and nasopharyngeal cancer
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EBV对胃癌、鼻咽癌WIF1、NLK、APC基因甲基化及表达的影响
DOI：
10.1002/jmv.24863
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发表时间：
2017-10-01
影响因子：
12.7
通讯作者：
Luo, Bing
中科院分区：
文献类型：
--
作者：
Zhao, Zhenzhen;Liu, Wen;Luo, Bing
EB病毒是一种重要的DNA肿瘤病毒，约10%的胃癌和99%的鼻咽癌与EB病毒有关。DNA甲基化和microRNAs(MiRNAs)是研究最多的表观遗传机制，可以促进疾病易感性。本研究旨在检测EBV对Wnt抑制因子1(WIF1)、Nemo样激酶(NLK)和结肠腺瘤性息肉病(APC)基因甲基化及在胃癌和鼻咽癌组织中表达的影响。用实时定量RT-PCR方法检测4株EBV阳性细胞和4株EBV阴性细胞中WIF1、NLK和APC基因的mRNA表达水平。用亚硫酸氢盐基因组测序或甲基化特异性聚合酶链式反应检测WIF1、NLK和APC启动子的甲基化状态。用5-氮杂胞苷(5-aza-DC)、miR-BART19-3p模拟物或抑制剂处理所有细胞系，用流式细胞仪和四甲基偶氮唑盐比色法检测细胞增殖情况。在所有8个细胞系中，WIF1、NLK和APC启动子都发生了高甲基化。5-氮杂-DC对细胞有生长抑制作用。转染miR-BART19-3p模拟物后，WIF1、APC表达降低，细胞增殖率增加。转染miR-BART19-3p抑制剂后，细胞表达水平升高，细胞生长受到抑制。在鼻咽癌和胃癌细胞系中，WIF1、NLK和APC的启动子高度甲基化，并且这三个基因的表达受miR-BART19-3p的调控。在EBV相关肿瘤中，miR-BART19-3p可增强Wnt通路的活性。
Epstein-Barr virus (EBV) is an important DNA tumor virus that is associated with approximately 10% of gastric carcinomas and 99% of nasopharyngeal cancers (NPC). DNA methylation and microRNAs (miRNAs) are the most studied epigenetic mechanisms that can prompt disease susceptibility. This study aimed to detect the effect of EBV on Wnt inhibitory factor 1 (WIF1), Nemo-like kinase (NLK), and adenomatous polyposis coli (APC) gene methylation, and expression in gastric carcinoma and NPC. The WIF1, NLK, and APC gene mRNA expression levels were measured by real-time quantitative RT-PCR in four EBV-positive cell lines and four EBV-negative cell lines. Bisulfite genomic sequencing or methylation-specific PCR was used to detect the methylation status of the WIF1, NLK, and APC promoters. All cell lines were treated with 5-azacytidine (5-aza-dC), miR-BART19-3p mimics or an inhibitor, and analyzed by flow cytometry and MTT cell proliferation assays. The WIF1, NLK, and APC promoters were hypermethylated in all eight cell lines. 5-Aza-dC displayed a growth inhibitory effect on cells. After transfection with miR-BART19-3p mimics, the expression of WIF1, and APC decreased, and the cellular proliferation rate increased. After transfection with the miR-BART19-3p inhibitor, the expression levels were higher, and the cell growth was inhibited. In the NPC and GC cell lines, the promoters of WIF1, NLK, and APC are highly methylated, and the expression of these three genes is regulated by miR-BART19-3p. The activity of the Wnt pathway in EBV-associated tumors may be enhanced by miR-BART19-3p.