Making a model of digestive tract infection and carcinogenisis of Epstein-Barr virus produced by epithelial cell line (GTC) derived from human gastric adenocarcinoma.

制作人胃腺癌上皮细胞系（GTC）产生的 Epstein-Barr 病毒的消化道感染和致癌模型。

• 批准号: 16591354
• 负责人: FUKUSHIMA Ryoji
• 金额: $ 2.3万
• 依托单位: Teikyo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591354/
• 关键词: Epstein-Barr virus; carcinogenisis; EBER; LMP-1; Balb; c mouse; EBV infection; 胃癌; EBV; マウス; EBウイルス; 消化器感染; マウス感染; EBV-NA; EBV-LHP-1; EBV-EBER; GTC-4-EBV

Epstein-Barr virus (EBV) has been implicated in the pathogenesis of some type of gastric adenocarcinoma. We established an epithelial cell line (GTC-4) derived form human gastric cancer tissue. The cell line have a capacity of producing EBV. We further cloned EBV producing cell line GTC-5 from GTC-4 cells and found that digestive tract of Balb/c mice can be infected by EBV produced by GTC-5 (GTC-5-EBV) and finally infected mice developed adenocarcinoma of the digestive tract.Balb/c mice were gavaged with 0.2ml of 50% ethanol to produce gastric mucosal injury and than GTC-5-EBV were gavaged. Histoimmunostaining showed that EBV nuclear antigen NA-2 were positive in the gastric mucosa 24 hours after the gavage of EBV and disappeared in 21 days. NA-1 and LMP-1 (latent membrane protein 1) were also present in gastric mucosa 5 days after the gavage and disappeared 6 months later. EBER (EBV-encoded small RNA) were identified in the gastric mucosa by in situ hybridization 24 hours after the gavage and it lasted for 9 months. EBER positive adenocarcinoma were developed in the stomach 9 months after the gavage of EBV.In the next experiment, Balb/c nude mice (6-7W and 10 month, n=10 each) were used to evaluate the carcinogenesis of GTC-5-EBV. GTC-5-EBV was injected weekly for 8 times. After 4 months, adenocarcinoma were found in the rectum of 100 % of elderly mice. In contrast, no cancer was found in the young mice during the observation period of 6 months.Our model of EBV associated carcinogenisis of digestive tract seems useful in studying the etiology and treatment of EBV associated disorders.

EB病毒（Epstein-Barr virus，EBV）与某些类型的胃腺癌的发病有关。我们从人胃癌组织中建立了一个上皮细胞系（GTC-4）。该细胞系具有产生EBV的能力。我们进一步从GTC-4细胞中克隆了EBV产生细胞系GTC-5，发现GTC-5-EBV可感染Balb/c小鼠的消化道，并最终导致消化道腺癌的发生。免疫组化染色显示，灌胃EBV后24 h胃粘膜中EBV核抗原NA-2阳性，21 d后消失。NA-1和LMP-1（潜伏膜蛋白1）也存在于胃粘膜管饲后5天，6个月后消失。胃粘膜原位杂交检测到EBER（EBV-encodedsmallRNA），EBER持续9个月。在第二次实验中，使用Balb/c裸小鼠（6- 7 W和10个月，每组n=10）来评估GTC-5-EBV的致癌性。GTC-5-EBV每周注射8次。4个月后，100%的老年小鼠直肠出现腺癌.而在6个月的观察期内，未发现幼龄小鼠患癌。我们建立的EBV相关消化道癌变模型对研究EBV相关疾病的病因和治疗是有用的。