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

• 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
• 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

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、肿瘤浸润白细胞的募集和激活、血管生成和转移，在癌症进展中起着至关重要的作用。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NF?B的调节。然而，我们实验室的研究表明，IL-8的转录调控不同于其他NF？B-dependent基因。具体来说，我们发现核I?B？在受刺激的白细胞中不抑制IL-8的表达，而抑制其他NF？B-dependent基因。此外，我们最近的数据显示，蛋白酶体抑制因其抑制NF？b依赖基因实际上增加了转移性前列腺癌和卵巢癌细胞以及人巨噬细胞中IL-8的表达。然而，其机制在很大程度上是未知的。提出的研究解决了缺乏对蛋白酶体抑制和核I?B?调控IL-8转录的认识。我们的长期目标是了解调节NF？B-responsive基因。本提案的目的是确定蛋白酶体和核I?B？调节IL-8的转录，这种调节与其他NF有何不同？B-dependent基因。中心假设是S536 p65磷酸化，特异性的DNA序列的NF？B结合位点和/或转录因子EGR-1使IL-8对I?B？，并增加IL-8的表达，以响应蛋白酶体的抑制。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Aim 1中，我们将验证以下假设：IL-8启动子主要被S536磷酸化的p65同型二聚体占据，蛋白酶体抑制S536的p65磷酸化，导致IL-8转录增加。在Aim 2中，我们将验证IL-8启动子序列和EGR-1参与是蛋白酶体抑制、IL-8表达增加和IL-8转录对I?B？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巨噬细胞作为两种Aims的模型。由于IL-8促进肿瘤细胞生长、血管生成、转移，激活白细胞，了解其在蛋白酶体和核I?B中的调控作用？可能在以IL-8过度表达为特征的癌症和炎症性疾病中具有重要的临床意义。此外，该项目将通过为有动力的贫困学生提供学习生物医学研究基础的大量机会来改善圣约翰大学的研究环境。

项目成果

Quantitative analysis of bortezomib-induced IL-8 gene expression in ovarian cancer cells.

硼替佐米诱导的卵巢癌细胞中 IL-8 基因表达的定量分析。

• DOI: 10.1007/978-1-4939-0928-5_27
• 发表时间: 2014
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Singha,Bipradeb;Phyo,SaiA;Gatla,HimavanthR;Vancurova,Ivana
• 通讯作者: Vancurova,Ivana

Anticancer drug bortezomib increases interleukin-8 expression in human monocytes.

• DOI: 10.1016/j.bbrc.2015.03.041
• 发表时间: 2015-05-01
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Sanacora, Shannon;Urdinez, Joaquin;Chang, Tzu-Pei;Vancurova, Ivana
• 通讯作者: Vancurova, Ivana

Transcriptional regulation of chemokine expression in ovarian cancer.

• DOI: 10.3390/biom5010223
• 发表时间: 2015-03-17
• 期刊: Biomolecules
• 影响因子: 5.5
• 作者: Singha B;Gatla HR;Vancurova I
• 通讯作者: Vancurova I

IKK inhibition increases bortezomib effectiveness in ovarian cancer.

IKK 抑制可增加硼替佐米治疗卵巢癌的疗效。

• DOI: 10.18632/oncotarget.4713
• 发表时间: 2015
• 期刊: Oncotarget
• 作者: Singha,Bipradeb;Gatla,HimavanthReddy;Phyo,Sai;Patel,Atish;Chen,Zhe-Sheng;Vancurova,Ivana
• 通讯作者: Vancurova,Ivana

Evaluating cytoplasmic and nuclear levels of inflammatory cytokines in cancer cells by western blotting.

通过蛋白质印迹评估癌细胞中炎症细胞因子的细胞质和细胞核水平。

• DOI: 10.1007/978-1-4939-0928-5_25
• 发表时间: 2014
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Gatla,HimavanthR;Singha,Bipradeb;Persaud,Valerie;Vancurova,Ivana
• 通讯作者: Vancurova,Ivana