Molecular basis of arsenic-induced cell transformation

砷诱导细胞转化的分子基础

• 批准号: 7012820
• 负责人: Zigang Dong
• 金额: $ 29.12万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7012820
• 关键词: JUN kinase; apoptosis; arsenic; athymic mouse; biological signal transduction; cell line; cell transformation; cell transplantation; chemical carcinogenesis; disease /disorder model; environmental contamination; enzyme activity; gene expression; histones; kinase inhibitor; mitogen activated protein kinase; molecular oncology; neoplastic transformation; phosphorylation; serine threonine protein kinase; small interfering RNA; transfection; western blottings

DESCRIPTION (provided by applicant): Arsenic is a well-documented human carcinogen. Our goal is to address the central hypothesis that MAP kinases, p90 S6 kinases, and histone H3 are mediators of arsenic-induced signal transduction and cell transformation. The Specific Aims are: Specific Aim 1, to determine whether arsenic-induced p38 kinase is required for arsenic-induced cell transformation; Specific Aim 2, to determine the role of c-Jun N-terminal kinases (JNKs) in arsenic-induced cell transformation and apoptosis; Specific Aim 3, to determine whether activation of p90 S6 kinase/MAPKAP-K1 (p90RSK) is involved in arsenic-induced signal transduction and cell transformation; and Specific Aim 4, to determine the role of p38 kinase, JNKs, and p90RSK in arsenic-induced phosphorylation of histone H3 and the role of phosphorylation of histone H3 in cell transformation. The strategy for Specific Aim 1 is to use p38 kinase inhibitors or dominant negative mutants or siRNA to inhibit p38 kinase activation. For Specific Aim 2, we will (1) test whether inactivation of JNK phosphorylation is required for development of resistant phenotype to arsenic; (2) test whether inhibition of JNK phosphorylation/activation will cause inhibition of arsenite-induced cell transformation or apoptosis; (3) test whether introduction of an active JNK mutant into arsenic resistant cells will cause the rescue of the apoptosis-resistant phenotype. For Specific Aim 3, we will inhibit p90RSK using cell lines expressing a dominant negative mutant or siRNA of p90RSK and cell lines which are deficient in p90RSK. For Specific Aim 4, we will (1) block the activation of JNK, p38, and p90RSK by specific inhibitors, dominant negative mutants, siRNA, or gene knockout cell lines; (2) use cell lines expressing siRNA of histone H3, dominant negative mutants (histone H3 S10 mutated to A10, H3 S28 mutated to A28) and histone H3 wild-type to study the role of histone H3 in arsenic-induced cell transformation. Such knowledge will facilitate the design of more effective and specific strategies with fewer side effects for chemoprevention of arsenic-induced cancer.

描述（由申请人提供）：砷是一种有据可查的人类致癌物。我们的目标是解决以下中心假设：MAP 激酶、p90 S6 激酶和组蛋白 H3 是砷诱导的信号转导和细胞转化的介质。具体目的是： 具体目的1、确定砷诱导的细胞转化是否需要砷诱导的p38激酶；具体目标2，确定c-Jun N末端激酶（JNK）在砷诱导的细胞转化和凋亡中的作用；具体目标3，确定p90 S6激酶/MAPKAP-K1（p90RSK）的激活是否参与砷诱导的信号转导和细胞转化；具体目标 4，确定 p38 激酶、JNK 和 p90RSK 在砷诱导的组蛋白 H3 磷酸化中的作用以及组蛋白 H3 磷酸化在细胞转化中的作用。具体目标1的策略是使用p38激酶抑制剂或显性失活突变体或siRNA来抑制p38激酶激活。对于具体目标 2，我们将 (1) 测试 JNK 磷酸化失活是否是砷抗性表型发展所必需的； (2)测试抑制JNK磷酸化/活化是否会抑制亚砷酸盐诱导的细胞转化或凋亡； (3)测试将活性JNK突变体引入砷抗性细胞中是否会引起抗凋亡表型的拯救。对于具体目标 3，我们将使用表达 p90RSK 显性失活突变体或 siRNA 的细胞系以及 p90RSK 缺陷的细胞系抑制 p90RSK。对于特定目标 4，我们将 (1) 通过特定抑制剂、显性失活突变体、siRNA 或基因敲除细胞系来阻断 JNK、p38 和 p90RSK 的激活； (2)利用表达组蛋白H3的siRNA、显性失活突变体(组蛋白H3 S10突变为A10、H3 S28突变为A28)和组蛋白H3野生型的细胞系来研究组蛋白H3在砷诱导的细胞转化中的作用。这些知识将有助于设计更有效、更具体、副作用更少的策略来化学预防砷诱发的癌症。