RESTRICTION LANDMARK GENOMIC ANALYSIS OF CANCER

描述(改编自《调查员摘要》)：本申请 重点介绍了利用限制性地标基因组扫描(RLGS)检测 有助于多步骤的遗传和表观遗传改变 肿瘤发生学。RLGS可以有效地检测基因扩增、基因丢失、 它的独特之处在于它能够检测DNA甲基化的变化。 分布在整个基因组上的大约2,000个基因座可以 在一次分析中被检测到。能够直接从 RLGS凝胶可以立即进入感兴趣的基因组区域。 建议利用两种小鼠肿瘤模型系统进行实验，肝脏 主要尿蛋白(MUP)/SV40T抗原转基因的致瘤性研究 小鼠与多发性肠道肿瘤的发生发展 (Min)在APC基因的小鼠同源基因中有突变的小鼠。这个 模型小鼠肿瘤系统允许某种程度上受控的肿瘤发生途径 在特定的遗传背景下，可以通过实验进行操作。 肿瘤相关的改变可以迅速定位于特定的染色体 精神错乱。实验将集中在鉴定和分离斑点克隆。 和其他与两个拷贝所在的遗传位点相对应的基因组克隆 在肿瘤发生过程中经常被删除和/或高甲基化。基因 在这些区域内将被识别以确定是否存在基因 改变(基因丢失、甲基化或扩增)改变基因功能 在肿瘤形成过程中。初步研究将集中在地区上 由两个点克隆S238鉴定，S238被发现对应于 P161NK4a和B330，识别15号染色体上的一个基因组区域 (S15Ncvs1)经历菌株和肿瘤特异性改变的 甲基化。这些研究应该为识别小说提供一种手段 参与肿瘤发生的基因，以及进一步了解 这一过程中的DNA甲基化。

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.