Generation of the wild-type Tsc2 transgenic Eker rat and its effect on renal carcinogenesis.

野生型 Tsc2 转基因 Eker 大鼠的产生及其对肾癌发生的影响。

基本信息

批准号：
08680915
负责人：
KOBAYASHI Toshiyuki
金额：
$ 1.54万
依托单位：
Japanese Foundation for Cancer Research
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08680915/
关键词：
Eker rat Hereditary tumoar Renal cancer Embryonic lethality Tsc2 gene Transgenic rat

项目摘要

We have identified a predisposing germline mutation for the Eker rat hereditary renal carcinoma in the rat homelog of human tuberous sclerosis 2 gene (TSC2). Also we have indentified somatic mutations and deletions on the wild-type Tsc2 allel (second hits) in renal carcinomas and even in preneoplastic lesions fom the Eker rat, indicating that the Tsc2 gene functions as a tumor suppressor. In tahis study, an attempt to suppress phenotypes of the Eker rat, the renal carcinogenesis in heterozygotes for Tsc2mutataion and the embryonic lethality in homozygous Tsc2 mutants, was carried out by introduction of a wild-type Tsc2 transgere. The transgene used here consisted of a wild-type rat Tsc2 cDNA (lacking exons 25 and 31 by alternative splicing events) and genomic DNA,and was driven by Tsc2 promoter. By idntroduction of this transgene, both of renalcarcinogenesis and embryonic lethality were successfully suppressed in the Eker rat. These results are final evidence to define the germline mutataion of Tsc2 gene as the cause of Eker rat phenotypes. Interesltingly, rescued homozygous Tsc2 mdutants developed renal tumors as well as pituitary tumors, which often developed in the Eker draats. In these tumors, the wild-type Tsc2 transgene was somatically deleted, indicating that the initiation of these tumors was caused by loss of Tsc2 function. the transgenic rescue for the Eker rat phenotype established in this study will be a usefull in vivo experimental system to analyze the function of Tsc2 gene and its product, and to elucidate molecular mechanism of tumorigenesis caused gy Tsc2 mdutations.

我们已经确定了人类结核性硬化症2基因（TSC2）大鼠Homelog中Eker大鼠遗传性肾癌的易感种系突变。同样，我们在肾癌中乃至肾脏癌的野生型TSC2 ALLEL（第二命中）的体细胞突变和缺失，甚至在eker大鼠的肿瘤肿瘤病变中，这表明TSC2基因起作用是肿瘤抑制剂。在塔希斯（Tahis）的研究中，通过引入野生型TSC2 Transgere的引入来执行一种尝试抑制Eker大鼠表型，TSC2Mutataion的肾脏癌变和TSC2Mutataion的肾脏癌变和纯合TSC2突变体的胚胎致死性。此处使用的转基因由野生型大鼠TSC2 cDNA（通过替代剪接事件缺少外显子25和31）和基因组DNA，并由TSC2启动子驱动。通过对这种转基因的研究，在Eker大鼠中成功抑制了肾癌发生和胚胎致死性。这些结果是将TSC2基因的种系变异构成Eker大鼠表型的原因的最终证据。相互繁殖的，救出的纯合TSC2 MDUTANT会形成肾脏肿瘤以及垂体肿瘤，这些肿瘤通常在Eker Draats中发展。在这些肿瘤中，野生型TSC2转基因被体形删除，表明这些肿瘤的启动是由TSC2功能丧失引起的。本研究中建立的Eker大鼠表型的转基因救援将是一个有用的体内实验系统，用于分析TSC2基因及其产物的功能，并阐明肿瘤发生的分子机制导致Gy TSC2 Mdutations。