CHROMOSOME BREAKPOINTS & RENAL & SMALL CELL LUNG CANCER

染色体断点

• 批准号: 2092865
• 负责人: David I Smith
• 金额: $ 24.03万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 1996-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2092865
• 关键词: antineoplastic antibiotics; aphidicolin; chromosome deletion; chromosome translocation; chromosome walking; cytogenetics; gene rearrangement; genetic library; genetic manipulation; genetic mapping; genetic markers; human genetic material tag; hybrid cells; kidney neoplasms; molecular cloning; molecular oncology; neoplasm /cancer genetics; northern blottings; nucleic acid hybridization; nucleic acid sequence; polymerase chain reaction; pulsed field gel electrophoresis; small cell lung cancer; southern blotting; tissue /cell culture

Specific rearrangements of human chromosome 3 are characteristically associated with certain malignant disorders. Loss of heterozygosity studies have revealed a consistent deletion of 3p2l.1 in spontaneous carcinoma of the kidney. Cytogenetic studies of studies of several hundred small cell and non-small cell lung cancer cell lines reveal three distinct chromosome 3 regions consistently deleted; 3pl3-l4.2, 3p2l and 3p24.2-pter. Chromosome 3pl4.2 is also the location of the most common constitutive fragile site which may be involved in the pathogenesis of some of these rearrangements. The large size of 3pl3-p2l precludes identifying all genes in this region and testing them individually for their role in tumor development. In the present application we propose to build upon the already constructed physical map to clone and characterize several chromosome breakpoints that may represent a short-cut to identifying .genes involved in cancer development. The first is a t(3;6)(p2l.l;pl3) breakpoint associated with hematologic malignancies. We have already isolated a cosmid which spans this breakpoint and contains a gene apparently disrupted by the translocation. We will isolate and characterize this gene and all other genes in a 200 kb region around this breakpoint. The second is the t(3;8)(pl4.2;ql4.13) breakpoint associated with hereditary renal cell carcinoma. We have already isolated markers which flank this breakpoint and propose to walk from the closest proximal marker (less than 500 kb) using overlapping YAC clones. We will then characterize this region and all genes within several hundred kb of the breakpoint. Finally we plan to extend our studies on the analysis of aphidicolin-induced fragile site breaks that occur in the 3pl3-l4.2 region. Using a sensitive PCR-based assay we will screen large numbers of chromosome 3-only somatic cell hybrids in which chromosome breakage is induced with aphidicolin. This will show whether there is a clustering of fragile site breaks. We will then use the closest flanking probes as startpoints for chromosome walking to this region. The entire region will be isolated in overlapping YAC clones and tested to determine if sequences from this region can induce chromosome fragility in an in vivo system. In addition to elucidating the structure of the most common fragile site in the genome this project may also lead to the identification of genes that are consistently deleted in small cell lung and renal cell carcinoma.

人类3号染色体的特异性重排 与某些恶性疾病有关 杂合性缺失 研究已经揭示了自发性肿瘤中3p21.1的一致缺失， 肾癌 几项研究的细胞遗传学研究 数百个小细胞和非小细胞肺癌细胞系揭示了三个 不同的3号染色体区域一致缺失; 3 p13 - 14.2、3 p2 l和3 p2 l。 3p24.2-pter. 染色体3p14.2也是最常见的 组成性脆性部位，可能参与的发病机制 其中一些重组。 3 p13-p2 l的大尺寸排除了鉴定该基因组中的所有基因。 区域，并单独测试它们在肿瘤发展中的作用。 在本申请中，我们提议在已经的基础上进行构建 构建物理图谱，克隆和表征多条染色体 断裂点可能代表识别相关基因的捷径 在癌症发展中的作用 第一个是t（3;6）（p21.1; pl 3）断点 与血液恶性肿瘤相关。 我们已经分离出一个 粘粒跨越这个断点，显然含有一个基因， 被易位破坏了 我们将分离并描述 基因和所有其他基因在这个断点周围的200 kb区域。 的 第二个是与遗传相关的t（3;8）（pl4.2;ql4.13）断点 肾癌 我们已经分离出了 断点，并建议从最近的近端标记（小于 超过500 kb）。 然后我们将描述 该区域和断裂点几百kb内的所有基因。 最后，我们计划将我们的研究扩展到分析 蚜虫素诱导的脆性位点断裂发生在3 p13 - 14.2 地区 使用灵敏的PCR检测，我们将筛选大量 3号染色体只有体细胞杂交，其中染色体断裂是 用aphidicolin诱导。 这将显示是否存在聚类 脆弱部位的断裂 然后我们将使用最近的侧翼探针， 染色体步移到这个区域的起始点。 整个地区 将在重叠的YAC克隆中分离，并进行测试以确定 来自该区域的序列可以在体内诱导染色体脆性， 系统 除了阐明最常见的 基因组中的脆弱位点，该项目也可能导致 鉴定小细胞肺中一致缺失的基因 和肾细胞癌。