Molecular biologic and pathologic study of Epstein-Barr virus infected lymphepithalial carcinomas in salivary gland

EB病毒感染唾液腺淋巴上皮癌的分子生物学和病理学研究

基本信息

批准号：
12576024
负责人：
SAKU Takashi
金额：
$ 9.73万
依托单位：
NIIGATA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12576024/
关键词：
lymphoepithelial carcinoma salivary gland tumor EBV LMP1 gene gene polymorphism cell culture PCR carcinogenetic mutations DNAシークエンス LMP-1 30bp欠損 in-situハイブリダイゼーション中国ロシア

项目摘要

A total 180 cases of salivary gland lymphoepithelial carcinoma (LEC) were col lected from Russia-Asian countries. Their paraffin blocks of surgical materials were obtained for imunohistochemistry, in-situ hybridization and DNA extractionImmunohistochemically, about 50% of the cases showed positive signals for LMP1 gene products, and 100% of them had EBER-1 RNA signals in tumor cells' nuclei, which suggested a definite EBV infection among salivary LEC cases. From 40 cases of them, the C-terminus of the LMP1 gene was PCR-amplified and direct-sequenced. The results indicated that there were at least two different strains of EBV harboring in salivary LEC and that there were many mutational events in them including the characteristic 30 by delefion. From 12 cases of them, the complete sequence for, the LMP1 gene were obtained, and many and new mutations in addition to those which were already found in nasapharyngeal LEC. The promoter region of the LMP1 gene was also analyzed from 20 cases o … More f them, and all of the cases shared common mutational events in regions which were expected to be associated with transcription factorsOne of the complete sequence obtained as mentioned above were cloned into an expression vector and the constructs were transfected into 293T cells to determine their effectd on the activation of NFkB pathways as well as cell growth. The transfected cells showed significant elevation of NFkB activities and suppressed cell growth, which were also seen in cells transfected with vectors with nasopharyngeal CAO or prototype B95-8. The results indicated that the expression of LMP1 affects transcritption activities in LEC cells but that the overall mutaions found in the present study were not always reasons for the tumorigenesisPrimary cultures for salivary LECs were tried many times. However, the cells were hard to maintain. One of them are growing now and they were characterized by immunofluorescence and PCR to be epithelial and EBV infectedThe results obtained as above has given us a new insight for EBV roles in the pathogenesis of salivary LEC. To this end, it is necessary to carry out further investigations on molecular functions of EBV oncogenes Less

总共有180例唾液腺淋巴上皮癌（LEC）从俄罗斯 - 亚洲国家（Imuno-Hervials）中获得了imuno-Hextemith，现场杂交和DNA提取性杂交，并获得了约50％的LMP1基因产品，并获得了imuno-Histosochemith。它们在肿瘤细胞的核ROM 40病例中具有EBER-1 RNA信号，LMP1基因的C端被PCR扩增并直接延迟。在其中的12个病例中，有许多特征，包括12例的压缩序列，LMP1基因，以及许多是Alleyngeal Lec的Thosse的新变异。 f他们，所有案例在惠勒（Whele）中共有共同的突变，预计将与序列化的ASBOVE序列的因子相关，并将构建体转染到293T细胞中，以确定对NFKB途径激活的影响。以及细胞的生长。转染的NFKB活性显着，并抑制了鼻cho的CAO或原型B95-8。尝试了许多液体，以维持一个一个矿石。对EBV毒素分子功能的研究较少