Usp16-Mediated Histone H2A Deubiquitination Regulates Breast Cancer Cell Invasion

Usp16 介导的组蛋白 H2A 去泛素化调节乳腺癌细胞侵袭

• 批准号: 9888396
• 负责人: Junjun Liu
• 金额: $ 10.32万
• 依托单位: CALIFORNIA STATE POLY U POMONA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9888396
• 关键词: Affect; BMI1 gene; Biography; Breast Cancer Cell; Breast Cancer Model; Cancer Patient; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; Cell Differentiation process; Cells; Cessation of life; Data; Deubiquitination; Development; ERBB2 gene; Epigenetic Process; Epithelial; Epithelial Cells; Epithelium; Estrogen receptor positive; Event; Gene Expression; Goals; Histone H2A; Investigation; Knowledge; Lysine; Mediating; Mesenchymal; Metastatic breast cancer; Molecular; Monoubiquitination; Mus; Neoplasm Metastasis; Outcome Study; PLK1 gene; PRC1 Protein; Peptide Hydrolases; Phosphorylation; Phosphorylation Site; Play; Prevention; Primary Neoplasm; Principal Investigator; Process; Reagent; Regulation; Reporting; Research; Rest; Role; Site; TWIST1 gene; Testing; Tissues; Ubiquitin; Ubiquitination; Woman; cancer cell; epithelial to mesenchymal transition; experience; experimental study; gene repression; histone modification; knock-down; malignant breast neoplasm; member; migration; novel; novel strategies; overexpression; tumor; tumorigenesis; ubiquitinated H2A

PROJECT SUMMARY/ABSTRACT Many important cellular events are regulated by antagonistic mechanisms, and understanding how factors involved in the regulation of tumorigenesis execute their antagonistic functions is particularly important. Histone H2A mono-ubiquitination at K119 mediated by polycomb repressive complex 1 (PRC1) is an important epigenetic mark associated with epithelial-mesenchymal transition (EMT) and cell invasion during early metastasis. However, less is known about the function of histone H2A deubiquitination during these processes. Our long-term research goal is to elucidate the molecular mechanism underlying the regulation of histone H2A ubiquitination and deubiquitination, and determine the role of this histone modification in the regulation of cell cycle and tumorigenesis. To this end, the goal of the proposed research is to study the function of histone H2A deubiquitination mediated by ubiquitin specific peptidase 16 (USP16) in the regulation of EMT and invasion of breast cancer cells. Our preliminary data has shown that BMI1, a member of PRC1, promotes EMT and invasion of breast cancer cells. Since USP16 specifically deubiquitinates H2A at K119, the central hypothesis of this application is that the USP16 antagonizes the function of BMI1 and inhibits the EMT and invasion of breast cancer cells. Furthermore, we have previously identified USP16 as a novel substrate of polo-like kinase 1 (PLK1). Therefore, the role of PLK1 in USP16-mediated regulation of EMT and cell invasion will also be determined. The rationale for this hypothesis rests on the following: once the role of USP16-mediated H2A deubiquitination in the regulation of EMT and invasion of cancer cells is established, a novel tumorigenesis regulatory mechanism that involves both the ubiquitination and deubiquitination of H2A can be defined. The central hypothesis will be tested through the activities including: (1) Determine whether USP16-mediated deubiquitination of histone H2A inhibits EMT and invasion of breast cancer cells. Following overexpression and knockdown of USP16, the changes in the expression of EMT markers and invasiveness of breast cancer cells will be analyzed. (2) Determine the role of PLK1 in USP16-mediated regulation of EMT and invasion of breast cancer cells. As the phosphorylation sites on USP16 have been identified, we will utilize both non- phosphorylatable and phosphomimic USP16 constructs to carefully evaluate the function of USP16 phosphorylation by PLK1 in regulating EMT and cell invasion. The expected outcomes of this study include the identification of ubiquitination and deubiquitination of histone H2A as an important mechanism for the regulation of metastasis in breast cancer, and the revealing of a cell cycle regulation-independent function of PLK1 in tumorigenesis. Knowledge of these mechanisms may contribute to the development of novel approaches in the prevention and treatment of metastatic breast cancer. !

项目总结/摘要 许多重要的细胞事件都是由拮抗机制调节的，了解这些因素如何影响细胞的生长， 参与肿瘤发生的调控，执行其拮抗功能尤为重要。组蛋白 多梳抑制复合物1（PRC 1）介导的K119上H2 A单泛素化是一种重要的 与上皮-间质转化（EMT）和早期细胞侵袭相关的表观遗传标记 转移然而，很少有人知道组蛋白H2 A去泛素化在这些过程中的功能。 我们的长期研究目标是阐明组蛋白H2 A调控的分子机制 泛素化和去泛素化，并确定这种组蛋白修饰在细胞调节中的作用。 周期和肿瘤发生。为此，本研究的目标是研究组蛋白H2 A的功能 泛素特异性肽酶16（USP 16）介导的去泛素化在EMT和侵袭中的调节作用 乳腺癌细胞我们的初步数据表明，BMI 1，PRC 1的成员，促进EMT， 乳腺癌细胞的侵袭。由于USP 16特异性地在K119处去泛素化H2 A，因此中心假设 USP 16拮抗BMI 1的功能并抑制EMT和侵袭， 乳腺癌细胞此外，我们以前已经确定USP 16作为polo样激酶的新底物， 1（PLK1）。因此，PLK 1在USP 16介导的EMT和细胞侵袭调节中的作用也将被进一步研究。 测定该假设的基本原理基于以下内容：一旦USP 16介导的H2 A的作用 去泛素化在调节EMT和癌细胞侵袭中的作用已经建立，这是一种新的肿瘤发生机制。 可以定义涉及H2 A的泛素化和去泛素化的调节机制。的 中心假设将通过以下活动进行检验：（1）确定USP 16介导的 组蛋白H2 A的去泛素化抑制乳腺癌细胞的EMT和侵袭。在过度表达后， USP 16基因的敲低、EMT标志物表达的变化和乳腺癌细胞侵袭力的变化 将被分析。(2)确定PLK 1在USP 16介导的EMT调节和乳腺浸润中的作用 癌细胞由于USP 16上的磷酸化位点已经被鉴定，我们将利用非磷酸化位点和非磷酸化位点。 可磷酸化的和拟磷酸化的USP 16构建体，以仔细评估USP 16的功能 PLK 1磷酸化在调节EMT和细胞侵袭中的作用。本研究的预期成果包括： 鉴定组蛋白H2 A的泛素化和去泛素化作为一个重要的机制， 调节乳腺癌转移，并揭示了细胞周期调节独立的功能， PLK 1与肿瘤发生这些机制的知识可能有助于开发新的 转移性乳腺癌的预防和治疗方法。 !