Arsenic and Nickel Carcinogenesis in Human Lung Cells

砷和镍对人肺细胞的致癌作用

• 批准号: 10245059
• 负责人: Max Costa
• 金额: $ 40.1万
• 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/10245059
• 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; lung cancer cell; mRNA Stability; member; migration; mimetics; neurodevelopment; osteoblast differentiation; overexpression; p38 Mitogen Activated Protein Kinase; prevent; protein expression; transcription factor; tumor; tumorigenesis; 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; lung cancer cell; 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.

项目摘要 SATB2表达对于金属癌变至关重要。 SATB2是唯一上调的常见基因 BEAS2B细胞由正常Beas2b细胞中的各种致癌金属和SATB2敲低的转化 防止金属癌变。在砷（AS）或镍（Ni）转化的细胞中敲除SATB2，导致 转型标志的损失，包括锚定独立生长，增强的细胞侵袭和 迁移。 SATB2的调节涉及miR-31和miR23a。 runx2转录因子下调 抑制SATB2表达并增加runx2的miRNA通过暴露于Ni和AS带来的miRNA 通过减弱这些翻译抑制性miR的表达来间接诱导SATB2。机制 暴露于AS和Ni后，Runx2的增加表达，在AS或Ni转化的细胞中 将被研究。 Runx2激活似乎是一个关键的上游事件，它增加了SATB2蛋白 保持癌症标志。 runx2将在正常的BEAS2B中被淘汰，以研究其在SATB2表达中的作用 以及由Ni和AS引起的细胞转化。 miR-31和miRNA 23A的Runx2和Antogomers将是 过表达和评估了BEAS2B转换的发生率。这些后来的实验将解决 这些miRNA如何阻止SATB2 mRNA的翻译。我们将利用Runx2的化学抑制剂（A1-10-- 104）研究是否由Ni和AS和AS诱导的细胞转化，通过过表达Runx2转化的细胞 被抑制。 A1-10-104也将用于研究其对裸鼠Xenographing肿瘤形成的影响。我们 将使用runx2芯片seq识别被AS和NI处理的单元格中的下游靶 金属以及用于过表达Runx2的单元格。 SATB2芯片seq将调查如何 转录因子维持癌症标志。我们将研究样品中的runx2/miRNA/satb2轴 人类肺癌和周围正常组织的。我们将研究NI和接触的机制 诱导BEAS2B细胞转化，增加和维持高水平的Runx2 mRNA和蛋白质以及 维护激活Runx2所需的翻译后修改。如何 RUNX2将下调miRNA-31和23A。我们将研究mir-的给药 慢病毒载体中的Mimetics（miRNA-31OR 23A）可以收缩BEES2B诱导的原位或异种肿瘤 与具有AS，Ni或过表达的SATB2相比，与具有A549的肺癌细胞相比 SATB2的低水平。