Analysis of AND-34 in human breast cancer

AND-34 在人类乳腺癌中的分析

• 批准号: 7076885
• 负责人: ADAM LERNER
• 金额: $ 23.66万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7076885
• 关键词: breast neoplasms; cell line; cell surface receptors; clinical research; drug resistance; estrogen inhibitor; focal adhesion kinase; growth factor receptors; guanine nucleotide exchange factors; hormone regulation /control mechanism; hormone sensitivity /resistance; hormone therapy; human tissue; immunocytochemistry; neoplasm /cancer genetics; phosphatidylinositol 3 kinase; posttranslational modifications; small interfering RNA; western blottings

DESCRIPTION (provided by applicant): The mechanisms by which human breast carcinomas acquire resistance to antiestrogen therapy remain incompletely understood. In an unbiased search for genes which regulate this process, Dorssers et al determined that upregulation of either a novel protein, BCAR3, or the focal adhesion adapter protein p130Cas are reproducible genetic events that induce such antiestrogen resistance. In concurrent and independent work, we cloned murine AND-34/BCAR3 and demonstrated that it associates with p130Cas through a Cdc25 GEF-like domain. Using Western analysis, we find that AND-34 is expressed at high levels in a subset of human breast carcinomas but not normal breast tissue. We subsequently determined that overexpression of AND-34 induces PI3K, Akt and Rac activation in breast cancer cell lines. Specific Aims: 1) To determine the mechanism by which overexpression of AND-34 in human breast cancer cells induces PI3K activation, we will: A) Examine whether AND-34 associates with and positively regulates PI3K activation by transmembrane tyrosine kinase receptors in breast cancer cell lines. This work will include an analysis of signaling in AND-34 -/- tissues; B) Examine the role of p130Cas in AND-34-mediated PI3K activation; C) Examine the role of Src kinases in such PI3K activation. 2) To determine how AND-34 and related gene family members affect PI3K signaling and clinical outcome in primary human breast cancers and to examine the effect of AND-34 on ER signaling, we will: A) Contrast the ability of human NSP1, AND-34 and NSP-3 to induce Akt and Rac activation and antiestrogen resistance in human breast cancer cell lines. B) Determine by Western analysis whether NSP1, AND-34 or NSP-3 expression in primary human breast cancer specimens correlates with PI3K activation or ERa expression; C) Determine by immunohistochemistry whether NSP1, AND-34 or NSP-3 expression in primary human breast cancer specimens correlates with ERa expression, disease-free or overall survival. D) Determine if AND-34 overexpression induces ligand-independent ERa-mediated ERE transactivation.

描述（由申请人提供）：人类乳腺癌获得抗雌激素治疗耐药性的机制仍不完全清楚。在对调节这一过程的基因的公正搜索中，Dorssers 等人确定，一种新蛋白 BCAR3 或粘着斑接头蛋白 p130Cas 的上调是可重复的遗传事件，可诱导此类抗雌激素耐药性。在并行和独立的工作中，我们克隆了小鼠 AND-34/BCAR3，并证明它通过 Cdc25 GEF 样结构域与 p130Cas 关联。使用 Western 分析，我们发现 AND-34 高水平表达 存在于人类乳腺癌的一个子集中，但不存在于正常乳腺组织中。我们随后确定 AND-34 的过度表达会诱导乳腺癌细胞系中的 PI3K、Akt 和 Rac 激活。具体目标： 1) 为了确定人乳腺癌细胞中 AND-34 的过度表达诱导 PI3K 激活的机制，我们将： A) 检查 AND-34 是否与乳腺癌细胞系中的跨膜酪氨酸激酶受体相关并正向调节 PI3K 激活。这项工作将包括对 AND-34 -/- 组织中的信号传导进行分析； B) 检查 p130Cas 在 AND-34 介导的 PI3K 激活中的作用； C) 检查 Src 激酶在此类 PI3K 激活中的作用。 2) 确定AND-34如何与相关 基因家族成员影响原发性人类乳腺癌的 PI3K 信号传导和临床结果 为了检查 AND-34 对 ER 信号传导的影响，我们将： A) 对比人 NSP1、AND-34 和 NSP-3 诱导人 Akt 和 Rac 激活以及抗雌激素抵抗的能力 乳腺癌细胞系。 B)通过Western分析确定原发性人乳腺癌标本中NSP1、AND-34或NSP-3的表达是否与PI3K激活或ERa表达相关； C) 通过免疫组织化学确定 NSP1、AND-34 或 NSP-3 是否表达 原发性人类乳腺癌样本中的 ERa 表达与无病生存或总生存相关。 D) 确定 AND-34 过度表达是否诱导配体独立的 ERa 介导的 ERE 反式激活。