SATB2 and Nickel Carcingenesis

SATB2 和镍致癌

• 批准号: 8998027
• 负责人: Max Costa
• 金额: $ 45.72万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8998027
• 关键词: A/J Mouse; AT Rich Sequence; Address; Animals; Archives; Arsenic; Automobile Driving; Binding; Breathing; Carcinogens; Cell Line; Cell Nucleus; Cells; Chromatin; Chronic; Cytoskeleton; DNA; Development; Dose; Embryo; Enzymes; Epigenetic Process; Epithelial Cells; Exposure to; Gene Activation; Gene Chips; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Gene Silencing; Gene Targeting; Genes; Grant; Health; Histones; Homeobox; Human; Ingestion; Ions; Lamins; Lung; Lung Neoplasms; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Mass Spectrum Analysis; Messenger RNA; Metal Carcinogenesis; Metals; MicroRNAs; Molecular; National Toxicology Program; Nickel; Nickel Subsulfide; Nose; Nuclear Envelope; Nuclear Matrix; Nuclear Matrix-Associated Proteins; Nuclear Pore; Nuclear Proteins; Oxides; Paper; Process; Property; Proteins; Rattus; Recruitment Activity; Role; Shapes; Small Interfering RNA; Structure; Sulfides; Testing; Tissues; Transformed Cell Line; Upstream Enhancer; Vanadates; Water; cancer cell; cancer risk; carcinogenesis; cell transformation; chromium hexavalent ion; gel electrophoresis; histone acetyltransferase; in vivo; knock-down; overexpression; promoter; small hairpin RNA; transcription factor; tumorigenesis

DESCRIPTION (provided by applicant): SATB2 is a homobox transcription factor first discovered in 2003 that binds to AT rich sequences and likely binds chromatin modifying enzymes such as histone acetyltranferases and deacetylases for the regulation of gene expression. A number of recent papers have described SATB2 overexpression in a variety of cancers and have emphasized the importance of this overexpressed gene in driving tumorigenesis. We have studied the malignant transformation of normal human bronchial epithelial cells (BEAS2B) by carcinogenic metals such as nickel (Ni), hexavalent chromium (Cr+6), arsenic (As) and vanadate (V). While each of these metals has its own unique signature of gene expression when they transform BEAS2B cells, the signature is vastly different from metal to metal. However, SATB2 is increased in every transformed clone by any one of these metals. SATB2 is not expressed in parental BEAS2B cells. Further studies have shown that SATB2 mRNA and protein are induced in BEAS2B cells by chronic Ni ion treatment. We hypothesize that SATB2 is a transcription factor needed for normal mammalian development but its inappropriate expression during chronic Ni exposure is a driver of cell transformation. We want to investigate the mechanisms and consequences of its overexpression in BEAS2B and 16HB cells exposed to and transformed by Ni. We will overexpress SATB2 in normal BEAS-2B and 16HBE cells and investigate the effect this has on the cell's transformed properties and study the expression of other genes in these cells using gene chips. SATB2 overexpression and resulting cell transformation will also be studied in the presence of nickel exposure. We will lower the levels of SATB2 by transient knockdown with siRNA and stable knockdown with small hairpin RNA in nickel transformed BEAS2B and 16HBE cells and study the consequences of SATB2 loss on their transformed properties and investigate the effect this knockdown has on the expression of other genes using gene chips in the nickel transformed cells. We will study the mechanism of SATB2 overexpression focusing on its promoter, enhancer, upstream regulators and miRNA that target SATB2 following chronic nickel treatment of BEAS2B and 16HBE cells and in nickel transformed cells. In addition, SATB2 overexpressed in BEAS2B and 16HBE cells and in Ni-transformed cells will be immunoprecipitated, and interacting protein partners will be identified by gel electrophoresis and mass spectrometry. To address whether nickel is able to induce SATB2 expression in vivo, we will expose A/J mice to various doses of nickel by inhalation or ingestion, and analyze SATB2 expression in several target tissues. To explore the role of SATB2 in nickel-induced tumorigenesis, we will analyze the levels of SATB2 expression in rat lung tumors induced by nickel subsulfide exposure (obtained from National Toxicology Program archive).

描述（由申请人提供）：SATB2是2003年首次发现的同源框转录因子，它与富含AT的序列结合，并可能结合染色质修饰酶，例如组蛋白乙酰转移酶和脱乙酰酶，以调节基因表达。最近的许多论文描述了 SATB2 在多种癌症中的过度表达，并强调了这种过度表达的基因在驱动肿瘤发生中的重要性。我们研究了镍（Ni）、六价铬（Cr+6）、砷（As）和钒酸盐（V）等致癌金属对正常人支气管上皮细胞（BEAS2B）的恶性转化。虽然每种金属在转化 BEAS2B 细胞时都有其独特的基因表达特征，但不同金属的特征有很大不同。然而，在每个转化克隆中，这些金属中的任何一种都会增加 SATB2。 SATB2 在亲代 BEAS2B 细胞中不表达。进一步的研究表明，长期镍离子处理可诱导 BEAS2B 细胞中 SATB2 mRNA 和蛋白的表达。我们假设 SATB2 是正常哺乳动物发育所需的转录因子，但其在慢性 Ni 暴露期间的不适当表达是细胞转化的驱动因素。我们想要研究其在暴露于 Ni 并被 Ni 转化的 BEAS2B 和 16HB 细胞中过度表达的机制和后果。我们将在正常 BEAS-2B 和 16HBE 细胞中过度表达 SATB2，并研究这对细胞转化特性的影响，并使用基因芯片研究这些细胞中其他基因的表达。 SATB2 过度表达和由此产生的细胞转化也将在镍暴露的情况下进行研究。我们将在镍转化的 BEAS2B 和 16HBE 细胞中通过 siRNA 瞬时敲低和小发夹 RNA 稳定敲低来降低 SATB2 水平，并研究 SATB2 缺失对其转化特性的影响，并研究这种敲低对其他基因表达的影响在镍转化细胞中使用基因芯片。我们将研究对 BEAS2B 和 16HBE 细胞以及镍转化细胞进行长期镍处理后 SATB2 过表达的机制，重点关注其启动子、增强子、上游调节子和靶向 SATB2 的 miRNA。此外，将免疫沉淀 BEAS2B 和 16HBE 细胞以及 Ni 转化细胞中过表达的 SATB2，并通过凝胶电泳和质谱法鉴定相互作用的蛋白伴侣。为了确定镍是否能够在体内诱导 SATB2 表达，我们将通过吸入或摄入将 A/J 小鼠暴露于不同剂量的镍，并分析几个靶组织中 SATB2 的表达。为了探讨 SATB2 在镍诱导的肿瘤发生中的作用，我们将分析亚硫化镍暴露诱导的大鼠肺肿瘤中 SATB2 的表达水平（从国家毒理学计划档案中获得）。