Somatic mutation of the tuberous sclerosis (Tsc2) tumor suppressor gene in chemically induced rat renal carcinoma cell
Somatic mutation of the tuberous sclerosis (Tsc2) tumor suppressor gene in chemically induced rat renal carcinoma cell
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DOI：
10.1097/00005392-199707000-00085
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发表时间：
1997-07-01
影响因子：
6.6
通讯作者：
Hino, O
中科院分区：
文献类型：
--
作者：
Urakami, S;Tokuzen, R;Hino, O
目的：在非遗传性散发性人肾细胞癌（RC）中检测到高频率的von Hippel-Lindau（VHL）基因突变。我们最近在Eker大鼠模型中鉴定了人类结节性硬化症（TSC）易感RC基因的大鼠同源物中的种系突变，并且在本研究中，我们在化学诱导的非Eker大鼠RC中寻找Tsc 2基因的突变。材料和方法：化学[N-乙基-N-羟乙基亚硝胺（EHEN）]诱导的非Eker大鼠RC系用PCR-单链构象多态性（PCR-SSCP）方法分析了BP 13和BP 36 B两株Tsc 2基因的41个编码外显子和1个非编码外显子。结果：BP 36 B基因第35外显子的迁移率发生了异常改变，与正常大鼠相比，BP 36 B基因第35外显子的迁移率发生了异常改变，与正常大鼠相比，BP 36 B基因第35外显子的迁移率发生了异常改变。该突变经直接测序证实为第1470位密码子第二位的T → C转换，导致氨基酸由亮氨酸变为脯氨酸（错义突变）。结论：这是首次证实非Eker大鼠肾细胞中存在Tsc 2基因体细胞突变。我们目前的研究结果呼吁注意进一步调查Tsc 2基因突变在大鼠肾癌发生中的作用和可能的Tsc 2基因突变在人类RC，特别是非透明细胞型，这是不相关的VHL基因。
Purpose: von Hippel-Lindau (VHL) gene mutations are detected in noninherited, sporadic human renal cell carcinomas (RCs) at a high frequency. We recently identified a germline mutation in the rat homologue of the human tuberous sclerosis (TSC) predisposing RC gene in the Eker rat model, and in this study we searched for mutations of the Tsc2 gene in chemically induced non-Eker rat RCs.Materials and Methods: Chemically [N-ethyl-N-hydroxyethylnitrosamine (EHEN)]-induced non-Eker rat RC lines (designated as BP13 and BP36B) were subjected to PCR-single strand conformation polymorphism (PCR-SSCP) analysis using specific primers covering entire exons of Tsc2 gene (41 coding exons and one non-coding exon). We simultaneously searched for mutations of Vhl gene, a rat homologue of von Hippel-Lindau disease gene (VHL) as well as Tsc2 gene.Results: BP36B showed an abnormal mobility shift from the normal tissue of the same rat in exon 35 on analysis by PCR-SSCP. This mutation was confirmed by direct sequencing and found to be a T-to-C transition at the second position of codon 1470, resulting in an amino acid change from leucine to proline (missense mutation).Conclusions: This is the first demonstration of Tsc2 gene somatic mutation in non-Eker rat RCs. Our present findings call attention to further investigation of the role of Tsc2 gene mutations in rat renal carcinogenesis and possible Tsc2 gene mutations in human RCs, especially of the non-clear cell type, which are not related to the VHL gene.