Research for Finding the Receptor of Epstein-Barr Virus Infection in Oral Squamous Cell Carcinomas

寻找口腔鳞状细胞癌中 Epstein-Barr 病毒感染受体的研究

• 批准号: 13671970
• 负责人: OOBU Kazunari
• 金额: $ 1.6万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671970/
• 关键词: Epstein-Barr virus; infection receptor; oral squamous cell carcinoma; CD21; cell to cell contact; Epstein-Barr Virus

We reported high incidence of Epstein-Barr virus (EVB) infection on both primary lesion and cervical lymphnodes in oral squamous cell carcinoma (OCSS). In order to reveal a mechanism of EBV infection to OSCC cells, EBV positive Akata cell (given from department of virology, Hokkaido Univ.) and CD21 cDNA (given from Alabama Univ. USA) were prepared. Squamous cell carcinoma cell line expressing EBV receptor CD21 (SAS-CD21-2) was established. SAS cells were transfected with 1μg pLNX-CR2 which contained CD21 and neomycin resistant genes, and had previously coated with lipopolyamine. Selection of cells containing stably integrated DNA was accomplished in medium containing Geneticin for 2 weeks. SAS transfectants were analyzed by flow cytometry using the OKB7 antibody. After the 10th passage, CD21 molecule was detected at the cell surface by analyses of flow cytometry. Therefore, we concluded that this transfectant cell line was stable line. The features of the SAS-CD21-2 were a little smaller than the original SAS cells. Growth doubling time of culture cells was approximately 25 hours. These SAS-CD21-2 cells were easier infected than the parent SAS cells by EB virus with the cell-to-cell contact manner like a case of gastric carcinomas (reported in 47^<th> Annual Meeting of Japanese Society of Oral and Maxillofacial Surgeons. 1st Nov. 2002, Sapporo).

我们报告了口腔鳞状细胞癌（OCSS）原发灶和颈部淋巴结中EB病毒（EVB）的高感染率。为了揭示EBV感染OSCC细胞的机制，将EBV阳性Akata细胞（来自北海道大学病毒学部门）和CD 21 cDNA（由亚拉巴马大学提供）。建立了表达EBV受体CD 21的鳞状细胞癌细胞系（SAS-CD 21 -2）。用1μg pLNX-CR 2转染SAS细胞，pLNX-CR 2含有CD 21和新霉素抗性基因，并预先用脂多胺包被。在含有遗传霉素的培养基中持续2周完成含有稳定整合DNA的细胞的选择。使用OKB 7抗体通过流式细胞术分析SAS转染子。第10代后，流式细胞仪检测细胞表面有CD 21分子。因此，我们认为该转染细胞系是稳定的。SAS-CD 21 -2细胞的特征比原始SAS细胞略小。培养细胞的生长倍增时间约为25小时。这些SAS-CD 21 -2细胞比亲本SAS细胞更容易被EB病毒感染，其细胞与细胞接触方式类似于胃癌病例（报道于47届<th>日本口腔颌面外科学会年会。2002年11月1日，札幌）。