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: 7122838
• 负责人: Jonathan J. Li
• 金额: $ 31.94万
• 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 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7122838
• 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 和侵袭性散发性乳腺癌 (BC) 的一个标志特征是其肿瘤细胞中高度的染色体不稳定性 (CIN) 和非整倍性。然而，到目前为止，CIN 和非整倍体尚未与雌激素 (E) 作用直接相关。该提案的目标是确定 E 在独特但相关的动物肿瘤模型（E 诱导的仓鼠肾恶性肿瘤 (HTK)）中引发 CIN 和非整倍性的机制。具体目标 1. 确定在 E 诱导的 HTK 发育过程中表现出持续过度表达/扩增的细胞周期成分（细胞周期蛋白 E、D 家族、B1）和调节剂（MDM2、DHFR）。这些基因/蛋白质已被证明可在体外系统中引发 CIN，并且是 E 介导的 c-myc/MYC 过表达/扩增的下游靶标。 具体目标 2. 确定在早期 E 诱导的 HTK 肿瘤发生过程中是否检测到 CIN是由 cyclin.cdk 复合物和其他细胞周期调节剂与纯化的 HTK 中心体及其蛋白质结合产生的中心体扩增引起的。这些研究将为启动中心体扩增的机制提供证据，从而为 CIN 和非整倍性提供证据，其中这些 cyclin.cdk 复合物和某些细胞周期调节剂优先与特定中心体蛋白或同一中心体蛋白上的不同位点结合。另一个目标是确定 HTK 发育早期阶段中心体扩增的最早时间出现。具体目标 3. 确定中心体扩增（在 Sp. Aim 2 中执行的数据）是否发生在 CIN 和非整倍性之前或作为它们的结果。这是阐明 CIN 和 HTK 中非整倍性机制的关键问题。这一具体目标的主要目标是根据 E 治疗的持续时间和 HTK 病灶大小（体积）确定早期 HTK 位点中 CIN 的最早出现时间；使用相同的标准来最早出现中心体扩增。第二个目标是利用相同的技术以及从 HTK 位点的 LCMD 和 DOP-PCR 生成的 DNA 样本进行跨物种微阵列分析，鉴定在 HTK 发育的最早阶段参与 CIN 或生长优势的候选基因。分析。