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原发灶和颈部淋巴结中EB病毒(EVB)感染的高发病例。为了揭示EBV感染口腔鳞癌细胞的机制，对EBV阳性的AKATA细胞(北海道大学病毒学系提供)和CD21基因(来自阿拉巴马大学)。美国)都做好了准备。建立了表达EBV受体CD21的鳞癌细胞系(SAS-CD21-2)。将含有CD2 1和新霉素抗性基因的1μg pLNX-CR2基因导入SAS细胞，并用脂多胺包被。在含有Geneticin的培养液中选择含有稳定整合DNA的细胞2周。用OKB7抗体进行流式细胞仪分析。第10代后，流式细胞仪检测到CD21分子出现在细胞表面。因此，我们认为该转基因细胞系为稳定的细胞系。SAS-CD21-2细胞的形态特征比原始细胞略小。培养细胞生长倍增时间约为25h。这些SAS-CD21-2细胞比亲代SAS细胞更容易被EB病毒以细胞间接触的方式感染(见日本口腔颌面外科学会第47届年会报道)。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).