ANALYSIS OF A NOVEL DNA AMPLIFICATION UNIT IN SARCOMAS

细胞癌基因的扩增经常发生在几个人类 癌症，并且是固体中增加基因表达的重要机制。 肿瘤的 肿瘤细胞中扩增基因的鉴定已被证明是 这是一种了解癌症遗传变异的有用方法。 先前用于从扩增的DNA序列中分离探针的方法 依靠组织培养细胞限制肿瘤的范围， 研究并提出了体外人工制品的问题。 我们绕过了 通过将扩增序列的凝胶复性与 聚合酶链反应。 利用这种方法，一种新的DNA 人恶性纤维组织细胞瘤活检组织的扩增单位 已经被识别并部分克隆 该放大单元 来源于染色体12 q13 -14，这是一个通常涉及 在软组织肿瘤中的重排，并含有至少一个转录的 区域（指定为肉瘤扩增序列的SAS）。 拟定研究 旨在详细描述SAS放大单元的特征， 基因水平，以确定其中表达的所有基因 在携带SAS扩增的细胞中。 这将通过使用 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YAC重叠群 它将被用来绘制肉瘤的这个区域。 使用探头， 从YAC克隆内的基因，将表达基因的cDNA 分离并表征。 这些信息将为 SAS扩增在肉瘤中作用的功能分析 肿瘤发生

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.