REGULATION OF CELL GROWTH BY THE WILMS TUMOR GENE

Wilms 肿瘤基因对细胞生长的调节

• 批准号: 6376614
• 负责人: ZHAO-YI WANG
• 金额: $ 12.7万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6376614
• 关键词: 3T3 cells; Wilms' tumor; athymic mouse; cell differentiation; cell growth regulation; cell line; developmental genetics; gene expression; gene interaction; genetic promoter element; growth inhibitors; immunoprecipitation; in situ hybridization; neoplasm /cancer genetics; neoplastic cell; neoplastic transformation; northern blottings; protein structure function; regulatory gene; retinoblastoma protein; transcription factor; tumor suppressor genes; tumor suppressor proteins

DESCRIPTION: (Adapted from investigator's abstract) The Wilms' tumor suppressor gene, wt1, encodes a zinc finger transcription factor, WT1. A number of mutations to the wt1 gene have been identified in subsets of Wilms tumor, mesothelioma, ovarian tumor and acute myeloid leukemia, suggesting that dysfunction of WT1 may be important in many tumors. However, effectively nothing is known about the endogenous genes regulated by WT1, and thus the signaling pathways triggered by WT1 and how mutated WT1 influences these pathways to result in Wilms' tumor remain to be discovered. The long-term objective of the investigator's research is to characterize the normal functions of WT1 during development and the dysfunctions of mutated WT1 that lead to the genesis of Wilms' tumor. The investigators recently cloned a gene whose expression is over tenfold greater in cells expressing WT1 than in cells with undetectable levels of WT1. The gene was identified as retinoblastoma (Rb) suppressor associated protein (AP) (RbAp46), a nuclear protein that physically interacts with Rb. The investigators have recently cloned and expressed the full-length cDNA of RbAp46, and analyzed its effect on tumor cells. Remarkably, expression of exogenous RbAp46 suppressed growth in four out of four tumor cell lines, suggesting that RbAp46 itself has growth inhibitory activity. The investigators now plan to establish the positive correlation between the levels of WT1 and RbAp46 expression in different tumor cells and in mouse tissues from different stages of development using Northern and in situ hybridization analysis. They plan to analyze the promoter region of RbAp46 to determine whether WT1 directly regulates the expression of RbAp46. They plan to explore the possible roles of RbAp46 as a mediator of WT1 function by expressing exogenous RbAp46 or by down-regulating RbAp46 in cells. The investigators plan to probe the mechanisms by which RbAp46 functions as a growth inhibitor by analyzing its impact on the cell cycle and its influence on the ras signaling pathway. The investigators plan to perform structure and function analysis to identify the domain(s) of RbAp46 required for its function. The investigators plan to identify the proteins which interact with RbAp46 by co-immunoprecipitation and an in vivo GST capture assay and, if necessary, by a yeast two-hybrid strategy. It is hoped that these studies will lay the foundation of therapeutic intervention for the Wilms' tumor and other tumors as well.

描述：（根据调查员的摘要改编）Wilms的肿瘤 抑制器基因WT1编码锌指转录因子WT1。 一个 WT1基因的突变数已在Wilms的子集中鉴定出来 肿瘤，间皮瘤，卵巢肿瘤和急性髓样白血病，表明 WT1的功能障碍在许多肿瘤中可能很重要。 然而， 实际上，关于WT1调节的内源基因，没有什么了解 因此，WT1触发的信号通路以及如何突变WT1 影响这些导致Wilms肿瘤的途径仍有待发现。 研究者研究的长期目标是表征 WT1在开发过程中的正常功能和功能障碍 突变的WT1导致Wilms肿瘤的起源。 调查人员 最近克隆了一个基因，其表达在细胞中的表达越高超过十倍 表达WT1的WT1比具有无法检测到的WT1水平的细胞中。 该基因是 被确定为视网膜母细胞瘤（RB）抑制剂相关蛋白（AP） （RBAP46），一种与RB物理相互作用的核蛋白。这 研究人员最近克隆并表达了 RBAP46，并分析了其对肿瘤细胞的影响。 值得注意的是，表达 外源RBAP46抑制了四分之四的肿瘤细胞系的生长， 表明RBAP46本身具有生长抑制活性。 这 研究人员现在计划建立 WT1和RBAP46在不同肿瘤细胞和小鼠中的表达水平 使用北部和原位的开发阶段的组织 杂交分析。 他们计划分析RBAP46的启动子区域 确定WT1是否直接调节RBAP46的表达。 他们 计划探索RBAP46作为WT1功能的中介者的可能作用 通过表达外源RBAP46或通过下调细胞中的RBAP46。 这 研究人员计划探测RBAP46用作一个机制 通过分析其对细胞周期及其影响的影响，生长抑制剂 在RAS信号通路上。 调查人员计划执行结构 和功能分析以识别RBAP46所需的RBAP46的域 功能。 研究人员计划识别相互作用的蛋白质 通过共免疫沉淀和体内GST捕获测定法和，使用RBAP46， 如有必要，通过酵母两种杂交策略。 希望这些 研究将奠定Wilms治疗干预的基础 肿瘤和其他肿瘤。