RESTRICTION LANDMARK GENOMIC ANALYSIS OF CANCER

癌症的限制性标志基因组分析

• 批准号: 6376185
• 负责人: WILLIAM A HELD
• 金额: $ 27.1万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-04-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6376185
• 关键词: DNA methylation; RNase protection assay; gastrointestinal neoplasms; gene deletion mutation; gene expression; gene mutation; genetic markers; genetic strain; genetic techniques; genetically modified animals; genome; laboratory mouse; liver neoplasms; molecular cloning; natural gene amplification; neoplasm /cancer diagnosis; neoplasm /cancer genetics; neoplastic process; northern blottings; oncogenes; polymerase chain reaction; restriction mapping; southern blotting

DESCRIPTION (Adapted from the Investigator's Abstract): This application focuses on utilizing Restriction Landmark Genomic Scanning (RLGS) to detect genetic and epigenetic alterations which contribute to multi-step tumorigenesis. RLGS can effectively detect gene amplification, gene loss, and is unique in its ability to detect alterations in DNA methylation. Approximately 2,000 loci distributed across the entire genome can be detected in a single analysis. The ability to "spot clone" directly from the RLGS gel allows immediate access to the genomic regions of interest. Experiments are proposed to utilize two model mouse tumor systems, liver tumorigenesis in the Major Urinary Protein (MUP)/SV40 T antigen transgenic mice, and intestinal tumorigenesis in the Multiple Intestinal Neoplasia (Min) mouse which has a mutation in the murine homolog of the APC gene. The model mouse tumor systems allow a somewhat controlled tumorigenetic pathway in a defined genetic background which can be manipulated experimentally. Tumor related alterations can be rapidly localized to specific chromosomal loci. Experiments will concentrate on identifying and isolating spot clones and other genomic clones corresponding to genetic loci in which both copies are frequently deleted and/or hypermethylated during tumorigenesis. Genes within these regions will be identified to establish whether the genetic alteration (gene loss, methylation, or amplification) alters gene function during tumorigenesis. Initial studies will concentrate on regions identified by two spot clones, S238 which was found to correspond to p161NK4a and B330 which identifies a genomic region on chromosome 15 (S15Ncvs1) that undergoes strain and tumor specific alterations in methylation. These studies should provide a means for identifying novel genes involved in tumorigenesis and further our understanding of the role of DNA methylation in this process.

描述（改编自研究者摘要）：本申请 重点是利用限制性标志基因组扫描（RLGS）检测 遗传和表观遗传改变， 肿瘤发生 RLGS可以有效地检测基因扩增、基因丢失、 并且在检测DNA甲基化改变的能力上是独特的。 分布在整个基因组中的大约2，000个基因座可以 在一次分析中发现。 能够“点克隆”直接从 RLGS凝胶允许立即接近感兴趣的基因组区域。 实验提出利用两种模型小鼠肿瘤系统，肝 主要尿蛋白（MUP）/SV 40 T抗原转基因小鼠的肿瘤发生 小鼠，以及多发性肠肿瘤中的肠肿瘤发生 (Min)在APC基因的鼠同源物中具有突变的小鼠。 的 模型小鼠肿瘤系统允许某种程度上受控的肿瘤发生途径 在一个确定的遗传背景下，可以通过实验来操纵。 肿瘤相关的改变可以快速定位于特定的染色体 的位点 实验将集中于鉴定和分离斑点克隆 以及对应于其中两个拷贝都 在肿瘤发生过程中经常缺失和/或高度甲基化。 基因 在这些区域内将被确定，以确定是否遗传 改变（基因丢失、甲基化或扩增）改变基因功能 在肿瘤发生过程中。 初步研究将集中于区域 通过两个斑点克隆鉴定，S238被发现对应于 p161 NK 4a和B330，其识别染色体15上的基因组区域 （S15 Ncvs 1）在细胞内经历菌株和肿瘤特异性改变， 甲基化 这些研究应提供一种方法， 参与肿瘤发生的基因，并进一步了解 DNA甲基化在这个过程中