SEQUENCING TUMOR SUPPRESSOR LOCI

申请人的描述（Applicant's Description） 我们的目标是利用基因组测序和分析， 我们假设候选肿瘤抑制基因位于 我们发现在人类癌症中是同源缺失的。 我们将：a） 确定完整和准确的核苷酸序列的 发现在人类癌症中重复缺失的染色体区域， 特别强调乳腺癌; B）使用序列分析， 数据库搜索以从这些区域中找到候选基因; c） 确定验证肿瘤的完整内含子/外显子结构 从这些区域转录的抑制基因。 目前正在对两个区域进行测序： 10号染色体携带PTEN，一种肿瘤抑制基因， 在晚期乳腺癌、脑癌和前列腺癌中经常发生突变;以及 20号染色体上的一个区域，在胃肠道疾病中经常被删除， 肿瘤的 多个位点正在进行表征，包括 8号染色体上的两个位点，21号染色体上的一个，4号染色体上的一个， in a variety品种of cancers癌症. 我们希望能测序和分析 每年5个位点，每个位点长度约300-400 kb。 这项工作将补充冷泉正在进行的其他工作 Harbor Cancer Center使用RDA（代表性差异 分析）来鉴定缺失的基因座，使用外显子捕获方法， 鉴定来自这些基因座的候选基因的杂交选择及其用途 突变分析，以验证候选基因参与 临床癌症

DESCRIPTION (Applicant's Description) Our objective is to utilize genomic sequencing and analysis to identify candidate tumor suppressors that we presume reside within regions that we have found are homozygously deleted in human cancers. We shall: a) determine the complete and accurate nucleotide sequence of the chromosomal regions found to be repeatedly deleted in human cancers, with a special emphasis on breast cancer; b) use sequence analysis and data base searching to find candidate genes from these regions; c) determine the complete intron/exon structures for the validated tumor suppressor genes transcribed from these regions. Sequencing of two regions is currently in progress: the region on chromosome 10 that harbors PTEN, a tumor suppressor gene that is frequently mutated in advanced breast, brain and prostate cancers; and a region on chromosome 20 that is frequently deleted in gastrointestinal tumors. Multiple loci are in the process of characterization, including two sites on chromosome eight, one on 21, and one on 4 that are deleted in a variety of cancers. We expect to be sequencing and analyzing about 5 loci per year, each about 300-400 kb in length. This work will complement other work in progress at the Cold Spring Harbor Cancer Center that utilizes RDA (representational difference analysis) to identify deleted loci, uses methods of exon trapping and hybrid selection to identify candidate genes from these loci, and uses mutational analysis to validate the involvement of candidate genes in clinical cancer.