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: 6173166
• 负责人: ZHAO-YI WANG
• 金额: $ 12.11万
• 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/6173166
• 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：（改编自研究者的摘要）威尔姆斯氏瘤 抑制基因 wt1 编码锌指转录因子 WT1。 一个 已在 Wilms 子集中鉴定出 wt1 基因的突变数量 肿瘤、间皮瘤、卵巢肿瘤和急性髓系白血病，提示 WT1 的功能障碍可能在许多肿瘤中很重要。 然而， 实际上，人们对 WT1 调节的内源基因一无所知， 以及 WT1 触发的信号通路以及 WT1 是如何突变的 影响这些途径导致维尔姆斯氏瘤的因素仍有待发现。 研究者研究的长期目标是表征 WT1在发育过程中的正常功能和功能障碍 WT1 突变导致肾母细胞瘤的发生。 调查人员 最近克隆了一个基因，其在细胞中的表达量高出十倍以上 表达 WT1 的细胞与 WT1 水平不可检测的细胞相比。 该基因是 被鉴定为视网膜母细胞瘤 (Rb) 抑制相关蛋白 (AP) (RbAp46)，一种与 Rb 发生物理相互作用的核蛋白。这 研究人员最近克隆并表达了全长 cDNA RbAp46，并分析了其对肿瘤细胞的作用。 值得注意的是，表达 外源性 RbAp46 抑制了四种肿瘤细胞系中四种的生长， 表明RbAp46本身具有生长抑制活性。 这 研究人员现在计划建立两者之间的正相关关系 不同肿瘤细胞和小鼠中 WT1 和 RbAp46 的表达水平 使用 Northern 和原位分析来自不同发育阶段的组织 杂交分析。 他们计划分析 RbAp46 的启动子区域 以确定WT1是否直接调节RbAp46的表达。 他们 计划探索 RbAp46 作为 WT1 功能中介者的可能作用 通过表达外源 RbAp46 或下调细胞中的 RbAp46。 这 研究人员计划探究 RbAp46 作为 通过分析生长抑制剂对细胞周期的影响及其影响 位于 ras 信号通路上。 研究人员计划进行结构 和功能分析以确定 RbAp46 所需的结构域 功能。 研究人员计划鉴定相互作用的蛋白质 通过免疫共沉淀和体内 GST 捕获测定与 RbAp46 结合， 如有必要，通过酵母二杂交策略。 希望这些 研究将为威尔姆斯氏症的治疗干预奠定基础 肿瘤和其他肿瘤也是如此。