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.

项目摘要 SATB 2的表达是金属致癌的关键。SATB 2是唯一一个在哺乳动物中上调的常见基因。 不同致癌金属对BEAS 2B细胞的转化及正常BEAS 2B细胞中SATB 2的敲低 防止金属致癌。在砷（As）或镍（Ni）转化的细胞中敲低SATB 2， 转化标志的丧失，包括锚定非依赖性生长、增强的细胞侵袭和 迁移SATB 2的调控涉及miR-31和miR 23 a。RUNX 2转录因子下调 抑制SATB 2表达和暴露于Ni和As引起的RUNX 2增加的miRNA 通过减弱这些翻译抑制miR的表达间接诱导SATB 2。的机制 在砷和镍暴露后以及在砷或镍转化细胞中诱导的RUNX 2表达增加 将被研究。RUNX 2激活似乎是增加SATB 2蛋白的关键上游事件， 保持癌症特征。RUNX 2将在正常BEAS 2B中被敲除以研究其在SATB 2表达中的作用 对Ni和As诱导的细胞转化也有抑制作用。RUNX 2和miR-31和miRNA 23 a的同源异构体将被 过表达并评估BEAS 2B转化的发生率。这些后期实验将解决 这些小RNA是如何阻止SATB 2 mRNA的翻译的。我们将使用RUNX 2的化学抑制剂（A1-10- 104)研究Ni和As诱导的细胞转化以及过表达RUNX 2诱导的细胞转化 被压制了。A1-10-104也将用于研究其对裸鼠异种移植瘤形成的影响。我们 将使用RUNX 2 Chip-Seq来鉴定用As和Ni处理的细胞中的下游靶点，或通过这些处理转化的细胞中的下游靶点。 金属，以及在细胞工程过表达RUNX 2。SATB 2 Chip-Seq将研究这一点 转录因子维持着癌症的特征。我们将研究样本中的RUNX 2/miRNA/SATB 2轴 人类肺癌和周围正常组织。我们将研究镍和砷暴露的机制， 诱导BEAS 2B细胞转化，增加并维持高水平RUNX 2 mRNA和蛋白，以及 维持激活RUNX 2所需的翻译后修饰。如何的机制 将研究RUNX 2下调miRNA-31和23 a。我们将研究是否给予miR- 慢病毒载体中的模拟物（miRNA-31或23 a）可以缩小BEAS 2B诱导的原位或异种移植肿瘤 用As、Ni或用过表达SATB 2转化的细胞，与用过表达SATB 2转化的A549肺癌细胞相比， 低水平的SATB 2。