Abnormality of sugar/amino acid transport and ATP sensor in renal carcinogenesis

糖/氨基酸转运和ATP传感器异常在肾癌发生中的作用

基本信息

批准号：
16590256
负责人：
KOBAYASHI Toshiyuki
金额：
$ 2.3万
依托单位：
Juntendo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590256/
关键词：
Hereditary renal cancer Animal model Eker rat Nihon rat Tsc2 Bhd ATP Amino acid 糖

项目摘要

It has been reported that the product of Tsc2 gene, which is the causative gene for the hereditary renal carcinoma (RC) of the Eker rat, functions in nutrient- and/or energy-regulated signal transduction. We found that the product of Bhd gene, which is the causative gene for the hereditary RC of the Nihon rat, shows partial amino acid sequence similarity with the LST7 product of Saccharomyces cerevisiae in amino-terminal half. Yeast mutants of LST7 gene have been reported to show defects in membrane transport system. We found that LST7-disruptant yeasts were slightly more sensitive to rapamycin than wild-type yeasts. However no apparent difference was observed in degree of resistance against toxic amino acid analogues tested so far. Introduction of rat wild-type or carboxy-terminally truncated folliculin-expression plasmids into mutant yeasts did not restore normal rapamycin sensitivity. We are analyzing other phenotypes of LST7 mutant and effect of mammalian folliculin expression in m … More utant yeasts. For analyze folliculin function using mammalian cells, we established and analyzed Bhd deficient renal tumor cell lines (1B series) in which rat Bhd cDNA was expressed by using the tet-off system. Compared with control cells (1R series) introduced with empty vector, 1B cells exhibited no apparent growth suppression. Expression of transpoters such as Gluts did not significantly change. However, 1B and 1R cells showed different morphological phenotypes. At phase-contrast microscopic level, cell adhesion and spreading were differ each other. By co-immunoprecipitation assay, we preliminarily found that the non-muscle myosin was co-precipitated using anti-Bhd antibody in 1B cell-specific manner. One plausible hypothesis is that Bhd products regulates cytoskeletal components as well as membrane transport systems. Through such functions, Bhd may modulates nutrient- or energy-regulated signal transuduction. We will clarify the function of Bhd product and its relation with nutrient/energy-regulated signals as well as Tsc2 product. Less

据报道，TSC2基因的乘积是Eker大鼠的遗传性肾癌（RC）的真正基因，在营养和/或能量调节的信号转导中起作用。我们发现，BHD基因的产物是Nihon大鼠的遗传RC的真正基因，它显示了与氨基末端一半酿酒酵母LST7产物的部分氨基酸序列相似性。 LST7基因的酵母突变体对我们进行了大鼠野生型或羧基末端截断的毛囊表达质粒的其他表型，并未恢复正常的雷帕霉素敏感性。我们对LST7突变体的其他表型和哺乳动物卵泡蛋白表达在M…更多的植物酵母菌中的作用进行了分析。为了使用哺乳动物细胞分析卵泡蛋白功能，我们建立了和分析了BHD缺陷肾脏肿瘤细胞系（1B系列），其中使用TET-OFF系统表达了大鼠BHD cDNA。与用空载体引入的对照细胞（1R系列）相比，1B细胞暴露于没有明显的生长抑制。发射器的表达（例如过剩的）没有显着变化。但是，1B和1R细胞显示出不同的形态表型。在相互对比的微观水平上，细胞粘合剂和扩散彼此不同。通过共免疫沉淀测定，我们初步发现，非肌肉肌球蛋白是使用抗BHD抗体以1B细胞特异性方式共沉淀的。一个合理的假设是，BHD产品调节细胞骨架成分以及膜运输系统。通过此类功能，BHD可以调节营养或能量调节的信号迁移。我们将阐明BHD产品的功能及其与营养/能量调节的信号以及TSC2产品的关系。较少的