Genetic Analysis of DBC2

描述(申请人提供)：一个新的基因，DBC2，最近被分离出来，对DBC2的初步研究表明，它是一个潜在的肿瘤抑制基因。本研究的长期目标包括对DBC2进行遗传学分析以验证其作为肿瘤抑制基因的有效性；对DBC2进行功能分析以阐明DBC2的生理和病理作用；以及DBC2的肿瘤生理学研究为乳腺癌和肺癌的发展提供新的见解，本研究项目的目标是验证DBC2是肿瘤抑制基因的假设，具体目的有：1.我们将利用可诱导的基因表达系统在体外测试DBC2的功能。突变将被引入到预测的功能域中。野生型和突变型DBC2将被诱导成各种类型的细胞。表型和表达模式的变化将被分析。具体目的2.研究DBC2在肿瘤中的转录调控，研究DBC2在细胞系和原发肿瘤中的表达及其启动子区域的甲基化状态。特定的目标3。为进一步研究DBC2的细胞功能奠定基础。将产生针对DBC2的抗体，并将分析DBC2在各种组织中的生理表达。具体目的4.我们将使用胚胎干细胞技术来建立Dbc2缺陷小鼠模型，并分析这些小鼠对肿瘤的敏感性。

DESCRIPTION (provided by applicant): A novel gene, DBC2, was recently isolated and preliminary studies of DBC2 indicate that it is a potential candidate for a tumor suppressor. The long-term objectives include genetic analysis of DBC2 to validate it as a tumor suppressor; functional analysis of DBC2 to clarify physiological and pathological roles of DBC2; and studies of tumor physiology of DBC2 to provide new insights to breast and lung cancer development, The goal of this research project is to test the hypothesis that DBC2 is a tumor suppressor gene, Specific aims are: Specific Aim 1. We will utilize inducible gene expression systems to test the DBC2 function in vitro. Mutations will be introduced into predicted functional domains. Wild type and mutated DBC2 will be induced into various types of cells. Alterations of phenotypes and expression patterns will be analyzed. Specific Aim 2. Transcriptional regulation of DBC2 in cancer will be studied, Expression of DBC2 and the methylation status of the DBC2 promoter region will be investigated in cell lines and primary tumors. Specific Aim3. We will lay the foundations for studies on cellular functions of DBC2. Antibodies against DBC2 will be generated, Physiological expression of DBC2 in various tissues will be analyzed. Specific Aim 4. We will use embryonic stem (ES) cell technology to generate a Dbc2-deficient mouse model and analyze these mice for tumor susceptibility.