Arsenic and Nickel Carcinogenesis in Human Lung Cells

砷和镍对人肺细胞的致癌作用

• 批准号: 10004646
• 负责人: Max Costa
• 金额: $ 41.77万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10004646
• 关键词: A549; AT Rich Sequence; Address; Adult; Anchorage-Independent Growth; Arsenic; Attenuated; Automobile Driving; Binding; Binding Proteins; Bone Development; CRISPR/Cas technology; Carcinogens; Categories; Cell Nucleus; Cells; ChIP-seq; Chemicals; Chromatin Loop; Cytoskeleton; DNA; DNA Sequence; Development; Down-Regulation; Embryo; Embryonic Development; Epithelial Cells; Event; Exposure to; Genes; Glandular Cell; Grant; Homologous Gene; Human; In Vitro; Incidence; Knock-out; Lamins; Lead; Lentivirus Vector; Lower Gastrointestinal Tract; Lung; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Matrix Attachment Regions; Mediating; Messenger RNA; Metal Carcinogenesis; Metals; MicroRNAs; Nickel; Normal tissue morphology; Nuclear Envelope; Nuclear Matrix; Nuclear Matrix-Associated Proteins; Nuclear Pore; Nuclear Proteins; Nude Mice; Paper; Pathway interactions; Play; Post-Translational Protein Processing; Process; Protein Family; Proteins; Regulation; Role; Sampling; Shapes; Structure; Tissues; Translations; Up-Regulation; Work; cancer cell; cancer type; carcinogenesis; carcinogenicity; cell transformation; cellular engineering; craniofacial development; experimental study; falls; gene repression; inhibitor/antagonist; knock-down; mRNA Stability; member; migration; mimetics; neurodevelopment; osteoblast differentiation; overexpression; p38 Mitogen Activated Protein Kinase; prevent; protein expression; transcription factor; tumor; tumorigenesis

PROJECT SUMMARY SATB2 expression is critical for metal carcinogenesis. SATB2 is the only common gene that is upregulated in BEAS2B cells transformed by various carcinogenic metals and knock-down of SATB2 in normal BEAS2B cells prevents metal carcinogenesis. Knockdown SATB2 in arsenic (As) or nickel (Ni) transformed cells, results in the loss in hallmarks of transformation including anchorage independent growth, enhanced cell invasion and migration. The regulation of SATB2 involves miR-31 and miR23a. RUNX2 transcription factor down regulates the miRNAs that inhibit SATB2 expression and increases in RUNX2 brought about by exposure to Ni and As indirectly induces SATB2 by attenuating expression of these translation inhibitory miRs. The mechanisms for the increase expression of RUNX2 induced following exposure to As and Ni, and in As or Ni transformed cells will be studied. RUNX2 activation appears to be a critical upstream event that increases SATB2 protein which maintains cancer hallmarks. RUNX2 will be knocked out in normal BEAS2B to study its role in SATB2 expression and for cell transformation induced by Ni and As. RUNX2 and antogomers of miR-31 and miRNA 23a will be overexpressed and the incidence of BEAS2B transformation assessed. These later experiments will address how these miRNA block the translation of SATB2 mRNA. We will utilize a chemical inhibitor of RUNX2 (A1-10- 104) to study whether cell transformation induced by Ni and As and, cells transformed by overexpress RUNX2 is suppressed. A1-10-104 will also be used to study its effect on xenograph tumor formation in nude mice. We will use RUNX2 Chip-Seq to identify downstream targets in cells treated with As and Ni or transformed by these metals, as well as in cells engineered to overexpress RUNX2. SATB2 Chip-Seq will investigate how this transcription factor maintains cancer hallmarks. We will investigate the RUNX2/miRNA/SATB2 axis in samples of human lung cancers and surrounding normal tissue. We will study the mechanisms of how Ni and As exposure induce BEAS2B cell transformation, increase and maintain high levels of RUNX2 mRNA and protein as well as maintaining the post-translational modifications that are required to activate RUNX2. The mechanisms of how RUNX2 downregulates miRNA-31 and 23a will be investigated. We will study whether administration of miR- mimetics (miRNA-31or 23a) in lentivirus vectors, can shrink orthotopic or xenograph tumors induced by BEAS2B cells transformed with As, Ni or with overexpressed SATB2, compared to A549 lung cancer cells that have low levels of SATB2.

项目总结 SATB2的表达在金属癌变过程中起关键作用。SATB2是唯一一个上调的常见基因 多种致癌金属对BEAS2B细胞的转化及SATB2基因在正常BEAS2B细胞中的表达 防止金属致癌。在砷(As)或镍(Ni)转化的细胞中敲除SATB2，导致 转化特征的丧失，包括锚定独立生长，增强的细胞侵袭和 迁移。SATB2的调控涉及miR-31和miR23a。Runx2转录因子下调 镍和砷暴露引起的抑制SATB2表达和增加RUNX2的miRNAs 通过减弱这些翻译抑制miR的表达间接诱导SATB2。实现以下目标的机制 砷镍诱导的RUNX2在砷镍转化细胞中的表达 将会被研究。Runx2的激活似乎是一个关键的上游事件，它会增加SATB2蛋白， 保持癌症的特征。Runx2将在正常BEAS2B中被敲除以研究其在SATB2表达中的作用 镍、砷诱导的细胞转化作用。MiR-31和miRNA23a的Runx2和异构体 并对BEAS2B转化的发生率进行了评估。这些稍后的实验将解决 这些miRNA如何阻止SATB2 mRNA的翻译。我们将使用RUNX2的化学抑制剂(A1-10- 104)研究镍、砷诱导的细胞转化和过表达RUNX2转化的细胞 是被压制的。A1-10-104还将用于研究其对裸鼠异种移植瘤形成的影响。我们 将使用RUNX2芯片序列来识别用As和Ni处理的细胞或由它们转化的细胞中的下游靶点 金属，以及在过表达RUNX2的细胞中。SATB2 Chip-Seq将调查这是如何 转录因子保持癌症的特征。我们将在样本中研究RUNX2/miRNA/SATB2轴 人类肺癌及其周围的正常组织。我们将研究镍和砷暴露的机制 诱导BEAS2B细胞转化，增加并维持高水平的RUNX2 mRNA和蛋白以及 维护激活RUNX2所需的翻译后修改。如何实现的机制 Runx2下调miRNA-31和23a将被调查。我们将研究是否给予miR- 慢病毒载体中的模拟物(miRNA-31或23a)可以缩小BEAS2B诱导的原位或异种移植瘤 AS、Ni或高表达SATB2的转化细胞与A549肺癌细胞相比 低水平的SATB2。