Interleukin-8 Regulation by Proteasome and Nuclear IkB in Cancer and Inflammation

蛋白酶体和核 IkB 对癌症和炎症中白细胞介素 8 的调节

基本信息

批准号：
8426626
负责人：
Ivana Vancurova
金额：
$ 49.5万
依托单位：
ST. JOHN'S UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-03-01 至 2016-08-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8426626
关键词：
Address Anti-Inflammatory Agents Anti-inflammatory Apoptotic Binding Binding Sites Biomedical Research Cell Proliferation Cell Survival Cells Clinical DNA Binding DNA Sequence Data Disease Environment Future Genes Genetic Transcription Goals Human Inflammation Inflammatory Interleukin-8 Knowledge Laboratories Learning Leukocytes Malignant Neoplasms Malignant neoplasm of ovary Malignant neoplasm of prostate Metastatic Prostate Cancer Modeling Neoplasm Metastasis Nuclear Phosphorylation Proteasome Inhibition Regulation Research Resistance Role Specific qualifier value Specificity Students Testing Transcriptional Regulation Universities angiogenesis base cancer cell cancer therapy cancer type career cell growth chemokine cytokine graduate student macrophage migration multicatalytic endopeptidase complex neoplastic cell p65 promoter public health relevance research study response transcription factor tumor tumor progression undergraduate student

项目摘要

DESCRIPTION (provided by applicant): Interleukin-8 (IL-8) is an inflammatory chemokine that has a crucial role in cancer progression through its induction of tumor cell proliferation, recruitment and activation of tumor-infiltrating leukocytes, angiogenesis, and metastasis. The expression of IL-8, as well as expression of many other inflammatory cytokines is regulated at the level of transcription by NF?B. However, studies from our laboratory indicate that the transcriptional regulation of IL-8 differs from the regulation of other NF?B-dependent genes. Specifically, we have found that the nuclear I?B? does not inhibit IL-8 expression in stimulated leukocytes, while it inhibits expression of other NF?B-dependent genes. In addition, our recent data show that the proteasome inhibition that is used as an anti-cancer therapy for its ability to inhibit expression of NF?B-dependent genes actually increases IL-8 expression in metastatic prostate and ovarian cancer cells and in human macrophages. However, the responsible mechanisms are largely unknown. The proposed research addresses the lack of knowledge on the regulation of IL-8 transcription by the proteasome inhibition and by nuclear I?B?. Our long-term goal is to understand the transcriptional mechanisms regulating expression of NF?B-responsive genes. The objective of this proposal is to determine how the proteasome and nuclear I?B? regulate transcription of IL-8, and how this regulation differs from other NF?B-dependent genes. The central hypothesis is that S536 p65 phosphorylation, specificity of the DNA sequence of the NF?B binding site, and/or the transcription factor EGR-1 render IL-8 unresponsive to the inhibition by I?B?, and increase IL-8 expression in response to the proteasome inhibition. Based on our preliminary data, the project will test two mutually non-exclusive models. In Aim 1, we will test the hypothesis that the IL-8 promoter is occupied predominantly by p65 homodimers phosphorylated on S536, and that the proteasome inhibition increases S536 p65 phosphorylation, resulting in the increased IL-8 transcription. In Aim 2, we will test the hypothesis that the IL-8 promoter sequence and EGR-1 involvement are responsible for the proteasome inhibition increased IL-8 expression and resistance of IL-8 transcription to I?B? inhibition. We will use metastatic prostate cancer PC-3 cells, ovarian cancer OVCAR-3 cells and stimulated U937 macrophages as models in both Aims. Since IL-8 promotes tumor cell growth, angiogenesis, metastasis, as well as activates leukocytes, understanding its regulation by proteasome and nuclear I?B? may have important clinical implications in cancers and inflammatory disorders characterized by excessive IL-8 expression. In addition, this project will enhance the research environment at St. John's University by providing motivated underprivileged students with numerous opportunities to learn the fundamentals of biomedical research.

描述（由申请人提供）：白细胞介素 8 (IL-8) 是一种炎症趋化因子，通过诱导肿瘤细胞增殖、招募和激活肿瘤浸润白细胞、血管生成和转移，在癌症进展中发挥关键作用。 IL-8 的表达以及许多其他炎症细胞因子的表达在转录水平上受到 NFκB 的调节。然而，我们实验室的研究表明IL-8的转录调控与其他NFκB依赖性基因的调控不同。具体来说，我们发现核I?B?不抑制受刺激白细胞中 IL-8 的表达，但抑制其他 NF?B 依赖性基因的表达。此外，我们最近的数据表明，蛋白酶体抑制能够抑制 NFκB 依赖性基因的表达，因此被用作抗癌疗法，实际上会增加转移性前列腺癌细胞和卵巢癌细胞以及人类巨噬细胞中的 IL-8 表达。。然而，其具体机制尚不清楚。拟议的研究解决了对蛋白酶体抑制和核 I?B? 调节 IL-8 转录的了解的缺乏。我们的长期目标是了解调节 NF?B 反应基因表达的转录机制。该提案的目的是确定蛋白酶体和核 I?B？调节 IL-8 的转录，以及这种调节与其他 NF?B 依赖性基因的不同之处。中心假设是 S536 p65 磷酸化、NFκB 结合位点 DNA 序列的特异性和/或转录因子 EGR-1 使 IL-8 对 IκB? 的抑制无反应，并增加 IL-8表达响应蛋白酶体抑制。根据我们的初步数据，该项目将测试两个互不排斥的模型。在目标 1 中，我们将测试以下假设：IL-8 启动子主要被 S536 上磷酸化的 p65 同二聚体占据，并且蛋白酶体抑制会增加 S536 p65 磷酸化，从而导致 IL-8 转录增加。在目标 2 中，我们将检验这样的假设：IL-8 启动子序列和 EGR-1 参与导致蛋白酶体抑制增加 IL-8 表达以及 IL-8 转录对 I?B? 的抗性。抑制。我们将使用转移性前列腺癌 PC-3 细胞、卵巢癌 OVCAR-3 细胞和刺激的 U937 巨噬细胞作为这两个目标的模型。由于 IL-8 促进肿瘤细胞生长、血管生成、转移以及激活白细胞，因此了解其通过蛋白酶体和核 I?B? 的调节。可能对以 IL-8 过度表达为特征的癌症和炎症性疾病具有重要的临床意义。此外，该项目还将为有积极性的贫困学生提供大量学习生物医学研究基础知识的机会，从而改善圣约翰大学的研究环境。