Genetic effectors of TGF-beta induced growth and arrest.

TGF-β 的遗传效应子诱导生长和停滞。

• 批准号: 7537893
• 负责人: BEN H PARK
• 金额: $ 5.96万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7537893
• 关键词: Ablation; Binding Sites; Biological Assay; Breast; Breast Cancer Cell; Breast Cancer Treatment; CDKN1A gene; Cancer Etiology; Cancer cell line; Candidate Disease Gene; Cell Line; Cell Proliferation; Cells; Cessation of life; Complementary DNA; Coupled; Deletion Mutation; Development; Elements; Epithelial; Epithelial Cells; Excision; Exons; G1 Arrest; Gene Expression; Gene Expression Microarray Analysis; Gene Targeting; Genes; Genetic; Genetic Recombination; Genetic Screening; Growth; Human; Lead; Link; Malignant Neoplasms; Mediating; Mediator of activation protein; Microarray Analysis; Modeling; Molecular Profiling; Protein Overexpression; Protein Tyrosine Phosphatase; RNA Interference; Regulation; Reporter; Research Personnel; Resistance; Role; Signal Transduction; System; TGF Beta Signaling Pathway; Tissue Microarray; Tissues; Transforming Growth Factor beta; United States; Up-Regulation; Work; base; cell growth; chromatin immunoprecipitation; cytokine; genetic analysis; genetic effector; homologous recombination; knock-down; malignant breast neoplasm; novel; oncoprotein p21; promoter; research study; resistance mechanism; response; restoration; tumor growth

Breast Cancer is a leading cause of cancer death in the United States with ~200,000 new cases and ~40,000 deaths each year. A better understanding of the genetic effectors of tumor growth and suppression would enable the identification of new targets for therapy. The aims of this proposal are directed at identifying and validating key components of the TGF-beta signaling pathway that would lead to the development of specific therapies for the treatment of breast cancer. AIM 1: Analyzingthe role of cdc25A in TGF-beta mediated growth stimulation of breast epithelial cells. Our initial studies demonstrated that TGF- beta mediated growth stimulation correlated with an increased expression of the cdc25A tyrosine phosphatase gene in cells lacking p21. In this aim we will further analyze the ability of cdc25A to mediate TGF-beta induced cellular proliferation using RNA interference, promoter/mutational/ChIP assays, and p21 cDNA mutational studies, coupled with an analysis of human breast cancers using tissue microarrays. AIM 2: Identifying mediators of TGF-beta induced cellular proliferation. In Aim 2, we hypothesize that there are other integral genes whose upregulation is needed for TGF-beta induced growth stimulation. We will use microarray expression analysis to identify these genes using our previously characterized p21-/- cell lines. In order to validate that these genes are true mediators of TGF-beta stimulated cellular proliferation, RNA interference gene "knock down" experiments and targeted gene ablation via homologous recombination will be performed. AIM 3: Identifying genetic effectorsthat mediate the growth response to TGF-beta. In this aim, we will identify additional effectors of TGF-beta mediated growth inhibition using a functional genetic screen. We will combine exon trapping with genetic instability to isolate genes whose inactivation confers resistance to TGF-beta mediated growth suppression. Isolated genes will be verified and validated by restoration of the putative gene and correlated with immunohistochemical studies using tissue microarrays.

乳腺癌是美国癌症死亡的主要原因，新病例约有200,000例 每年约40,000人死亡。更好地了解肿瘤生长和抑制的遗传效应因子 将能够确定新的治疗靶标。该提议的目的是针对的 识别和验证TGF-beta信号通路的关键组件，这将导致 开发用于治疗乳腺癌的特定疗法。目标1：分析CDC25A的作用 在TGF-β介导的乳腺上皮细胞的生长刺激中。我们的最初研究表明TGF- β介导的生长刺激与Cdc25a酪氨酸的表达增加有关 缺乏p21的细胞中的磷酸酶基因。在此目标中，我们将进一步分析CDC25A进行调解的能力 TGF-β使用RNA干扰，启动子/突变/芯片测定和P21诱导细胞增殖 cDNA突变研究，再加上使用组织微阵列对人乳腺癌的分析。目的 2：识别TGF-β诱导的细胞增殖的介体。在AIM 2中，我们假设有 TGF-β诱导生长刺激需要上调的其他积分基因。我们将使用 微阵列表达分析使用我们先前表征的p21 - / - 细胞系识别这些基因。在 为了验证这些基因是TGF-β刺激细胞增殖的真正介体RNA 干扰基因“击倒”实验和通过同源重组的靶向基因消融将 被执行。 AIM 3：确定遗传效应介导对TGF-β的生长反应。在这个 目的，我们将使用功能遗传来确定TGF-β介导的生长抑制的其他效应因子 屏幕。我们将将外显子捕获与遗传不稳定性结合在一起，以分离出灭活的基因 对TGF-β介导的生长抑制的抗性。孤立的基因将通过 恢复推定基因，并与使用组织微阵列的免疫组织化学研究相关。