ANALYSIS OF A NOVEL DNA AMPLIFICATION UNIT IN SARCOMAS

肉瘤中新型 DNA 扩增单元的分析

基本信息

批准号：
3201111
负责人：
PAUL S. MELTZER
金额：
$ 20.94万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-05-01 至 1993-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3201111
关键词：
artificial chromosomes complementary DNA gene expression genetic library histiocytoma human tissue in situ hybridization molecular cloning natural gene amplification neoplasm /cancer genetics northern blottings nucleic acid sequence polymerase chain reaction

项目摘要

Amplification of cellular oncogenes occurs frequently in several human cancers and is an important mechanism of increased gene expression in solid tumors. Identification of amplified genes in tumor cells has proven to be a useful approach for understanding genetic alterations in cancer. Previous procedures for isolating probes from amplified DNA sequences have relied on tissue culture cells limiting the range of tumors which can be studied and raising questions of in vitro artifact. We have circumvented these problems by combining in gel renaturation of amplified sequences with the polymerase chain reaction. Using this approach, a novel DNA amplification unit from biopsies of human malignant fibrous histiocytoma has been identified and partially cloned. This amplification unit is derived from chromosome 12q13-14, a site commonly involved in rearrangements in soft tissue tumors, and contains at least one transcribed region (designated SAS for sarcoma amplified sequence). Proposed studies are intended to characterize the SAS amplification unit in detail at the genetic level in order to identify all genes within it which are expressed in cells carrying SAS amplification. This will be accomplished using genomic analysis techniques based on yeast artificial chromosome (YAC) cloning. A YAC contig will be established for the SAS amplification unit which will be used to map this region in sarcomas. Using probes derived from genes within the YAC clones, cDNAs for expressed genes will be isolated and characterized. This information will provide a basis for the functional analysis of the role of SAS amplification in sarcoma oncogenesis.

细胞癌基因的扩增经常发生在几个人中癌症是固体基因表达增加的重要机制肿瘤。鉴定肿瘤细胞中扩增基因已被证明是理解癌症遗传改变的有用方法。从扩增的DNA序列中分离探针的先前程序已有依靠组织培养细胞限制了肿瘤的范围研究并提出了体外文物的问题。我们已经绕过通过将放大序列的凝胶恢复性结合到与聚合酶链反应。使用这种方法，一种新颖的DNA 来自人类恶性纤维组织细胞瘤活检的放大单元已被识别并部分克隆。这个扩增单元是源自染色体12q13-14，一个通常涉及的地点软组织肿瘤中的重排，至少包含一个转录区域（肉瘤扩增序列的SAS）。拟议的研究旨在详细描述SAS扩增单元遗传水平以识别其内部的所有基因在带有SAS扩增的细胞中。这将使用基于酵母人工染色体（YAC）的基因组分析技术克隆。将为SAS扩增单元建立一个YAC重叠群它将用于肉瘤中的该区域。使用得出的探针从YAC克隆内的基因中，用于表达基因的CDNA将是孤立和特征。此信息将为 SAS扩增在肉瘤中的作用的功能分析肿瘤发生。