Genomic Instability and Etiology of Estrogen Oncogenesis

雌激素肿瘤发生的基因组不稳定性和病因学

• 批准号: 6793711
• 负责人: Jonathan J. Li
• 金额: $ 32.71万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6793711
• 关键词: aneuploidy; breast neoplasms; cell cycle proteins; centrosome; dihydrofolate reductase; disease /disorder etiology; disease /disorder model; estrogens; fluorescent in situ hybridization; functional /structural genomics; hamsters; hormone related neoplasm /cancer; immunocytochemistry; in situ hybridization; laser capture microdissection; microarray technology; neoplasm /cancer genetics; polymerase chain reaction; protooncogene; southern blotting

DESCRIPTION (provided by applicant): A hallmark characteristic of DCIS and invasive sporadic breast cancer (BC) is the high level of chromosomal instability (CIN) and aneuploidy in its tumor cells. However, CIN and aneuploidy have not been, until now, directly related to estrogen (E) action. The goal of this proposal is to determine the mechanism whereby E elicits CIN and aneuploidy in a unique, but relevant animal tumor model, the E-induced malignant tumors in the hamster kidney (HTK). SPECIFIC AIM 1. To determine the cell cycle components (cyclins E, D family, B1) and modulators (MDM2, DHFR) which exhibit sustained overexpression/amplification during E-induced HTK development. These genes/proteins have been shown to elicit CIN in in-vitro systems, and are downstream targets of c-myc/MYC overexpression/amplification mediated by E. SPECIFIC AIM 2. To determine whether the CIN detected during early E-induced HTK oncogenesis is caused by centrosome amplification resulting from the binding of cyclin.cdk complexes and other cell cycle modulators to purified HTK centrosomes and their proteins. These studies will provide evidence for a mechanism for initiating centrosome amplification, and hence CIN and aneuploidy in which these cyclin.cdk complexes and certain cell cycle modulators bind preferentially to specific centrosome proteins, or to different sites on the same centrosome protein. Another goal is to determine the earliest temporal appearance of centrosome amplification during early stages of HTK development. SPECIFIC AIM 3. To determine whether centrosome amplification (data performed in Sp. Aim 2) occurs before CIN and aneuploidy or as a consequence of them. This is a key issue in elucidating the mechanism of CIN and aneuploidy in HTKs. A major goal of this specific aim is to determine the earliest temporal appearance of CIN in early HTK loci, based on duration of E-treatment and HTK foci size (volume); using the same criteria for the earliest appearance of centrosome amplification. A second goal is to identify candidate genes that are involved in either CIN or growth advantage in the earliest stages of HTK development, employing the same techniques and DNA samples generated from LCMD and DOP-PCR of HTK loci to be subjected to cross-species mircoarray analysis.

描述（由申请人提供）：DCIS和侵入性零星乳腺癌（BC）的标志性特征是其肿瘤细胞中高水平的染色体不稳定性（CIN）和非整倍性。但是，到目前为止，CIN和非整倍性与雌激素（E）作用直接相关。该提案的目的是确定在独特但相关的动物肿瘤模型中引起CIN和非整倍性的机制，即HAMSTER肾脏（HTK）的电子诱导的恶性肿瘤。具体目的1。确定细胞周期成分（Cyclins E，D家族，B1）和调节剂（MDM2，DHFR），它们在电子诱导的HTK发育过程中表现出持续的过表达/扩增。这些基因/蛋白质已被证明在体外系统中引起了CIN，并且是E.特定目标2介导的C-Myc/MyC过表达/扩增的下游靶标。确定在E-e-e诱导的HTK早期癌症中检测到的CIN是否是由Cyclied Cyclients cyclin and Cyclin and Cyclin and Cyclin.cdecline and cyclin.cdecline and Cyclin.cdemplectors and Cyclin.cdecline and Cyclin.cdemplecters and Cyclin.cdemplecters and cyclin.c declin.c cyclin.c cyclin.c cyclin.c cyclin。蛋白质。这些研究将为启动中心体扩增的机制提供证据，因此CIN和非整倍性在其中这些细胞周期蛋白。CDK络合物和某些细胞周期调节剂优先与特定的中心体蛋白质或在同一中心体蛋白上的不同位点结合。另一个目标是确定在HTK开发的早期阶段，最早的中心体扩增的时间出现。特定目标3。确定中心体扩增（在sp。AIM2中执行的数据）是在CIN和非整倍性之前还是出现它们的结果。这是阐明HTKS中CIN和非整倍性机理的关键问题。该特定目的的主要目的是根据电子处理的持续时间和HTK焦点大小（体积）确定早期HTK基因座CIN的最早时间出现；使用相同的标准来最早出现中心体扩增。第二个目标是确定在HTK开发的最早阶段与CIN或生长优势相关的候选基因，采用了HTK基因座的LCMD和DOP-PCR产生的相同技术和DNA样品，以进行跨特征mircoarray分析。