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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.

细胞癌基因扩增在几个人类中频繁发生并且是固体细胞中基因表达增加的重要机制肿瘤。肿瘤细胞中扩增基因的鉴定已被证明是这是一种了解癌症基因变化的有用方法。以前从扩增的DNA序列中分离探针的程序已经依靠组织培养细胞限制了肿瘤的范围研究并提出了体外人工制品的问题。我们已经绕过了通过将扩增序列的凝胶复性与聚合酶链式反应。使用这种方法，一种新的DNA 人恶性纤维组织细胞瘤活检组织的扩增单位已被鉴定并部分克隆。这个放大单元是起源于染色体12q13-14，这是一个常见的软组织肿瘤中的重排，并包含至少一个转录区域(指定为肉瘤扩增序列的SA)。建议的研究意在详细描述SAS放大单元基因水平，以便识别其中所有表达的基因在携带SAS扩增的细胞中。这将通过以下方式完成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YAC重叠群这将被用来绘制肉瘤中的这一区域。使用派生的探针从YAC克隆中的基因来看，表达基因的cDNA将是与世隔绝的，有特点的。这些信息将为 SAS基因扩增在肉瘤中作用的功能分析致癌作用。