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

我们报道了口腔鳞状细胞癌（OCSS）原发性病变和颈部淋巴结的eb病毒（EVB）感染的高发率。为了揭示EBV感染OSCC细胞的机制，我们制备了EBV阳性Akata细胞（来自北海道大学病毒系）和CD21 cDNA（来自美国阿拉巴马大学）。建立表达EBV受体CD21的鳞状细胞癌细胞系（SAS-CD21-2）。用含有CD21和新霉素耐药基因的1μg pLNX-CR2转染SAS细胞，并预先包被脂多胺。在含有Geneticin的培养基中选择含有稳定整合DNA的细胞2周。用OKB7抗体流式细胞术分析SAS转染物。第10代后，流式细胞术检测细胞表面CD21分子。因此，我们认为该转染细胞系是稳定的细胞系。SAS- cd21 -2细胞的特征比原SAS细胞略小。培养细胞生长倍增时间约为25小时。这些SAS- cd21 -2细胞比亲本SAS细胞更容易被EB病毒感染，细胞间接触方式类似于胃癌病例（报告于47^< 1990年日本口腔颌面外科学会年会上，2002年11月1日，札幌）。

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).