Analysis of micrometastasis using GFP-expressed metastasis model of oral cancer and inhibition of metastasis by new anti-tumor drugs

GFP表达的口腔癌转移模型分析微转移及新型抗肿瘤药物抑制转移

基本信息

批准号：
13557178
负责人：
HAMAKAWA Hiroyuki
金额：
$ 8.9万
依托单位：
Ehime University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

We established a stably transfected cell line which expresses high levels of green fluorescent protein (GFP), thus permitting the detection and visualization of developing tumors and lymph node metastases after injection into nude mice. Cells of the human oral squamous carcinoma cell line (SAS-L1) were transfected with an expression vector containing a CDNA encoding humanized GFP and the neomycin resistance gene. A clone with stable high-level expression of GFP was selected in vitro using G418. To study metastasis formation, GFP-expressing cells were injected orthotopically into the tongue of nude mice. The resultant tumor growth in the tongue and micrometastases in the lymph nodes could be visualized by GFP fluorescence. Therefore a useful model has been developed for the study of oral cancer, firstly to understand the metastatic process and secondly for the evaluation of potential treatments.Second, we analyzed the expression of COX2 and checked the effect of COX2 inhibitor. The purpose of this study is to investigate the relationship between the radiation sensitivity and elevated level of COX-2. Radiation sensitivity of the eight oral SCC cell lines differed greatly in their response to radiation. Further, the level of the COX-2 expression correlated inversely with increased tumor radiation sensitivity. The similar significant association between the response to preoperative radiation therapy and COX-2 overexpression was observed in the oral SCC patients. In addition, treatment with a COX-2 selective inhibitor enhanced the radioresponse of HSC-2 cell, which constitutively expressed COX-2. These results suggested that COX-2 expression level correlates to radiation tolerance and the COX-2 selective inhibitor may be a potent enhancer for tumor radioresponse in oral SCC.

我们建立了稳定的转染细胞系，该细胞系表达高水平的绿色荧光蛋白（GFP），从而允许在注射到裸鼠后的肿瘤和淋巴结转移的检测和可视化。人口腔鳞状癌细胞系（SAS-L1）的细胞用包含编码人性化GFP和新霉素耐药基因的cDNA的表达载体转染。使用G418在体外选择具有稳定的GFP高级表达的克隆。为了研究转移的形成，将表达GFP的细胞原位注射到裸鼠的舌头中。 GFP荧光可以观察到舌头中所得的肿瘤生长和淋巴结中的微转移。因此，已经为口腔癌研究开发了一个有用的模型，首先是为了了解转移过程，其次是对潜在处理的评估。第二，我们分析了COX2的表达并检查了COX2抑制剂的效果。这项研究的目的是研究辐射敏感性与COX-2水平升高之间的关系。八个口腔SCC细胞系的辐射敏感性在其对辐射的反应方面有很大不同。此外，COX-2表达的水平与肿瘤辐射敏感性的增加相关。在口腔SCC患者中观察到对术前放射疗法的反应与COX-2过表达之间的相似显着关联。此外，用COX-2选择性抑制剂处理增强了HSC-2细胞的辐射响应，HSC-2细胞组成型表达COX-2。这些结果表明，COX-2表达水平与辐射公差相关，COX-2选择性抑制剂可能是口服SCC中肿瘤辐射响应的有效增强子。