ISOLATION OF A HUMAN 11P112 LIVER TUMOR SUPPRESSOR GENE

人 11P112 肝肿瘤抑制基因的分离

• 批准号: 6173823
• 负责人: WILLIAM B COLEMAN
• 金额: $ 18.61万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 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 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6173823
• 关键词: cell line; chromosomes; complementary DNA; gene expression; genetic library; genetic mapping; human genetic material tag; laboratory rat; liver neoplasms; molecular cloning; nucleic acid sequence; solution hybridization; transfection; tumor suppressor genes

Numerous studies suggest that inactivation or deletion of one or more tumor suppressor genes may represent early and/or necessary steps in the neoplastic transformation of liver. Furthermore, comparative chromosomal mapping studies suggest that the development of liver tumors in humans and rodents may share a common molecular mechanism that involves inactivation of analogous tumor suppressor genes. Based upon these observations, we have suggested that chromosome transfer studies utilizing human chromosomes and rat liver tumor cell lines may facilitate the facile localization, identification, and cloning of human genes responsible for tumor suppression in liver. To this end, we have established and characterized a model for the functional identification of human tumor suppressor genes in rat liver tumor cell lines. Human chromosome 11, which has been implicated in the pathogenesis of human liver tumors, was introduced into highly aggressive rat liver epithelial tumor cell lines using a microcell-mediated chromosome transfer technique. We have demonstrated that human chromosome 11 suppresses the tumorigenic potential of some rat liver tumor cell lines, providing direct evidence for the presence of a liver tumor suppressor gene on this human chromosome. Further, we have established a microsatellite- based map of the liver tumor suppressor region, localizing the gene(s) to a 950-kb segment of human 11p11.2-p12 and forming the basis for positional cloning of candidate tumor suppressor genes. The studies contained in this proposal seek to identify, isolate, and characterize the human 11p11.2-p12 liver tumor suppressor gene. The goals of this proposal are to (i) establish a PAC contig spanning the liver tumor suppressor region of human 11p11.2-p12, (ii) utilize the isolated PAC clones in the construction of a complete physical map of this chromosomal region, (iii) isolate candidate liver tumor suppressor genes based upon complementarity to PAC clones corresponding to this region, (iv) characterize candidate liver tumor suppressor genes by DNA sequence analysis, and (v) determine the ability of candidate genes to express tumor suppressor activity in transfected rate liver tumor cell lines.

大量研究表明，一个或多个基因的失活或缺失 肿瘤抑制基因可能代表肿瘤生长过程中的早期和/或必要步骤。 肝脏肿瘤转化。 此外，比较 染色体作图研究表明肝脏肿瘤的发生 人类和啮齿动物可能具有共同的分子机制 涉及类似肿瘤抑制基因的失活。 基于 根据这些观察结果，我们建议染色体转移研究 利用人类染色体和大鼠肝肿瘤细胞系可能 促进人类的轻松定位、识别和克隆 负责肝脏肿瘤抑制的基因。 为此，我们有 建立并描述了功能识别模型 大鼠肝肿瘤细胞系中人类肿瘤抑制基因的研究。 人类 11号染色体与人类的发病机制有关 肝脏肿瘤，被引入高度侵袭性的大鼠肝上皮细胞 使用微细胞介导的染色体转移的肿瘤细胞系 技术。 我们已经证明人类 11 号染色体抑制 一些大鼠肝肿瘤细胞系的致瘤潜力，提供 肝肿瘤抑制基因存在的直接证据 这条人类染色体。 此外，我们还建立了一颗微卫星—— 基于肝脏肿瘤抑制区域的图谱，定位基因 到人类 11p11.2-p12 的 950-kb 片段，并形成 候选抑癌基因的定位克隆。 研究 该提案中包含的内容旨在识别、隔离和表征 人类 11p11.2-p12 肝脏肿瘤抑制基因。 本次活动的目标 建议 (i) 建立跨越肝肿瘤的 PAC 重叠群 人类 11p11.2-p12 的抑制区域，(ii) 利用分离的 PAC 克隆在构建完整的物理图 染色体区域，(iii) 分离候选肝脏肿瘤抑制基因 基于与该区域对应的 PAC 克隆的互补性， (iv) 通过 DNA 序列表征候选肝脏肿瘤抑制基因 分析，以及（v）确定候选基因的表达能力 转染率肝肿瘤细胞系中的肿瘤抑制活性。