Genetic Analysis of DBC2

描述（由申请人提供）：一个新的基因，DBC2，最近被分离出来，DBC2的初步研究表明它是一种潜在的肿瘤抑制因子。长期目标包括对DBC2进行基因分析以验证其作为肿瘤抑制因子；分析DBC2的功能，阐明DBC2的生理病理作用；研究DBC2的肿瘤生理学，为乳腺癌和肺癌的发展提供新的见解，本研究项目的目的是验证DBC2是肿瘤抑制基因的假设，具体目的是：我们将利用诱导型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。野生型和突变型DBC2将被诱导成各种类型的细胞。表型和表达模式的改变将被分析。具体目标2。将研究DBC2在癌症中的转录调控，在细胞系和原发肿瘤中研究DBC2的表达和DBC2启动子区域的甲基化状态。具体Aim3。为进一步研究DBC2的细胞功能奠定基础。产生针对DBC2的抗体，分析DBC2在各组织中的生理表达。具体目标我们将使用胚胎干细胞技术生成dbc2缺陷小鼠模型，并分析这些小鼠的肿瘤易感性。

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.