The Analysis of Molecular Pass Way of Apoptosis Induction with Human Oral Squamous Cell Carcinoma Cell Lines Containing p53 Mutations

p53突变人口腔鳞癌细胞株诱导凋亡的分子途径分析

基本信息

批准号：
10470428
负责人：
SAKAI Eiki
金额：
$ 8.38万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

To achive the successful transduction of the CD80 gene into oral SCC, we produced a recombinant adenovirus in which human CD80 cDNA was cloned in the E1-deficient Advector. We infected these tumor cell lines with CD80-Ad and cell surface expression of CD80 was determined by flow cytometry after 48 hr. Efficient and high levels of cell surface expression was observed in oral SCC cell lines. This results demonstrate that the use of adenovirus vector enables the transduction of a molecule into oral SCC cell lines.We examined the effect of gene-transduction with murine 4-1 BB ligand (CD137L), which is one of the tumor necrosis facter (TNF) families. 4-1 BBL costimulatory signals preferentially induce CD8+ T cell proliferation and IFN-γ production. Administration of mAb against 4-1 BB eradicated established several murine tumors. We transduced of 4-1 BB ligand (CD137L) into a murine squamous cell carcinoma. Introduction of the 4-1 BBL gene efficiently elicited anti-tumor immune responses in syngenic mice. Our results suggested that introduction of 4-1 BBL may induce efficient T cell immune responses against tumors.Surgically resected oral SCCs (n=37) were investigated by immunohistochemistry and 29 cases were positive for p53. Forty three samples diagnosed as leukoplakia with epithelial dysplasia in the oral cavity were subjected to immunohistochemical staining of p53 protein. Among these lesions, 12 were developed into SCC during the clinical observation. Nine out of above 12 lesions showed p53 positive staining even before malignant transformation to SCC.It was suggested that the immunohistochenical analysis of p53 protein is suggested to be a useful diagnostic procedure for oral Leukoplakia which probably develop future into SCC.

为了实现将 CD80 基因口服成功转导至 SCC，我们生产了一种重组腺病毒，其中人 CD80 cDNA 克隆到 E1 缺陷的 Advector 中，我们用 CD80-Ad 感染这些肿瘤细胞系，并测定了 CD80 的细胞表面表达。 48小时后通过流式细胞术在口腔SCC细胞系中观察到高效且高水平的细胞表面表达。该结果表明，腺病毒载体的使用能够实现。将分子转导到口腔 SCC 细胞系中。我们检查了小鼠 4-1 BB 配体 (CD137L) 基因转导的效果，该配体是肿瘤坏死因子 (TNF) 家族之一，优先诱导 4-1 BBL 共刺激信号。 CD8+ T 细胞增殖和 IFN-γ 产生。针对 4-1 BB 的 mAb 根除已建立的几种小鼠肿瘤。将 4-1 BBL 基因（CD137L）导入小鼠鳞状细胞癌中，可在同基因小鼠中有效引发抗肿瘤免疫反应。对切除的口腔鳞状细胞癌（n=37）进行免疫组织化学检查，其中 29 例 p53 呈阳性，43 例被诊断为上皮性白斑。对口腔异型增生进行p53蛋白免疫组织化学染色，其中12个在临床观察过程中发展为SCC，其中9个在恶变为SCC之前就显示出p53阳性染色。 p53 蛋白的免疫组织化学分析被认为是口腔白斑的一种有用的诊断方法，口腔白斑未来可能发展为鳞状细胞癌。