Arsenic and Nickel Carcinogenesis in Human Lung Cells

砷和镍对人肺细胞的致癌作用

• 批准号: 10681242
• 负责人: Max Costa
• 金额: $ 39.3万
• 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/10681242
• 关键词: A549; AT Rich Sequence; Adult; Anchorage-Independent Growth; Arsenic; 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; Invaded; Knock-out; Lamins; 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; 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.

项目概要 SATB2 表达对于金属致癌至关重要。 SATB2 是唯一上调的常见基因 多种致癌金属转化的 BEAS2B 细胞以及正常 BEAS2B 细胞中 SATB2 的敲低 防止金属致癌。在砷 (As) 或镍 (Ni) 转化细胞中敲低 SATB2，会导致 转化标志的丧失，包括贴壁独立生长、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 暴露于 Ni 和 As 后抑制 SATB2 表达并增加 RUNX2 的 miRNA 通过减弱这些翻译抑制性 miR 的表达来间接诱导 SATB2。的机制 暴露于 As 和 Ni 后以及在 As 或 Ni 转化细胞中诱导的 RUNX2 表达增加 将被研究。 RUNX2 激活似乎是增加 SATB2 蛋白的关键上游事件， 保持癌症特征。将在正常BEAS2B中敲除RUNX2以研究其在SATB2表达中的作用 以及 Ni 和 As 诱导的细胞转化。 RUNX2 以及 miR-31 和 miRNA 23a 的异构体将 过度表达并评估 BEAS2B 转化的发生率。这些后续实验将解决 这些 miRNA 如何阻断 SATB2 mRNA 的翻译。我们将利用 RUNX2 的化学抑制剂 (A1-10- 104) 研究Ni和As是否诱导细胞转化以及过表达RUNX2转化的细胞 被压制。 A1-10-104也将用于研究其对裸鼠异种肿瘤形成的影响。我们 将使用 RUNX2 Chip-Seq 来识别经 As 和 Ni 处理或经这些转化的细胞中的下游靶标 金属，以及被设计为过度表达 RUNX2 的细胞。 SATB2 Chip-Seq 将研究如何 转录因子维持癌症特征。我们将研究样本中的 RUNX2/miRNA/SATB2 轴 人类肺癌和周围正常组织。我们将研究 Ni 和 As 暴露的机制 诱导 BEAS2B 细胞转化，增加并维持高水平的 RUNX2 mRNA 和蛋白质以及 维持激活 RUNX2 所需的翻译后修饰。其机制如何 将研究 RUNX2 下调 miRNA-31 和 23a。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诱导的原位或异种肿瘤 与使用 As、Ni 或过表达 SATB2 转化的细胞相比，A549 肺癌细胞具有 SATB2 水平低。