Inflammatory Cytokines Link NF-kappaB with Bim Degradation and Metastasis

炎症细胞因子将 NF-kappaB 与 Bim 降解和转移联系起来

基本信息

批准号：
8257586
负责人：
MARTY W MAYO
金额：
$ 27.61万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-01 至 2014-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8257586
关键词：
Address American Amino Acids Anoikis Apoptosis Apoptotic Binding Biological Assay Cancer Etiology Cell Death Cell Survival Cell model Cells Cessation of life Clinical Colon Carcinoma Complex Cues Cullin Proteins Data Diagnosis Diagnostic Neoplasm Staging Disease Epithelial Epithelial Cells Extracellular Matrix Frequencies Genetic Transcription Goals Growth Factor Half-Life Human In Vitro Inflammation Mediators Inflammatory Interleukin-1 Laboratories Ligands Link Lung Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of pancreas Malignant neoplasm of prostate Maps Mediating Mesenchymal Microtubules Mitochondria Molecular Morphology NF-kappa B Neoplasm Metastasis Newly Diagnosed Non-Small-Cell Lung Carcinoma Nuclear Outcome Pathway interactions Phenotype Phosphorylation Polyubiquitination Process Property Proteins Regulation Resistance Role Secondary to Site Small Interfering RNA Staging Staining method Stains Stimulus TNF gene Tumor stage Ubiquitination United States Up-Regulation Withdrawal Work Xenograft Model cancer cell cancer stem cell chemotherapy cytochrome c cytokine in vitro Model in vivo Model lung small cell carcinoma malignant breast neoplasm member mortality multicatalytic endopeptidase complex mutant novel novel strategies outcome forecast p65 prevent pro-apoptotic protein protein degradation protein expression receptor research study response self-renewal synaptotagmin I transcription factor tumor tumor growth ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): Lung cancer causes more mortality in the United States than any other form of cancer. Non-small lung cancer (NSCLC) is the most common form of the disease, constituting over 85% of all newly diagnosed lung cancer. The poor clinical prognosis for NSCLC is due to late stage diagnosis and high frequency of metastatic disease. NF-?B is an immediate-early transcription factor that is commonly up regulated in NSCLC tumors. NSCLC tumors display elevated NF-?B activity, which correlates with invasive de-differentiated morphology, tumor staging, and poor clinical outcomes. Many laboratories, including our own, have shown that NF-?B provides cellular resistance to many forms of apoptosis. To understand better how NSCLC cells resist apoptotic cues, we have characterized the molecular pathways used by NF-?B to block apoptosis. Preliminary data presented in this proposal indicate that the p65 component of NF-?B provides a novel avenue of cell survival by blocking detachment from the extracellular matrix. Pro inflammatory cytokines inhibit this form of apoptosis in a process referred to as anoikis. Both IKK and p65 activities are required to block anoikis by inhibiting Bim, a pro-apoptotic protein important for induction of anoikis in NSCLC cells. IKK and p65 are required to stimulate Bim protein turnover in response to pro-inflammatory cytokines. IKK¿ directly phosphorylates Bim, regulating its half-life through a process involving polyubiquitination and proteasome-dependent degradation. Knockdown of the adaptor molecule COMMD1, a member of ECSSOCS1 E3 ligase complex that regulates p65 degradation, results in a significant up regulation of endogenous Bim protein expression. Using an in vitro model for invasion and metastasis that induces epithelial-mesenchymal transition (EMT), we demonstrate that pro-inflammatory cytokines facilitate TGF¿-induced EMT. The EMT phenotype is associated with an elevation in IKK and p65 activity, loss of Bim expression, and resistance to anoikis. Since EMT has been proposed to link de-differentiation with the self-renewing properties observed in cancer stem cells, we hypothesize that loss of Bim protein expression is one of the rate-limiting steps in NSCLC metastasis. The goal of this proposal is to understand how p65 regulates Bim protein turnover and to determine whether this pro-inflammatory responsive mechanism governs NSCLC metastasis. To address our hypothesis three Specific Aims will be addressed. Aim 1 will determine whether stimulation or inactivation of IKK and p65 activities alters the sensitivity of NSCLC cells to anoikis. Experiments will identify the minimal interacting domain required to stimulate Bim degradation and will determine if localization of p65 to the microtubule complex or to the mitochondria alters Bim induced apoptosis. Aim 2 will determine the importance of the COMMD1-containing E3 ligase for ubiquitination of Bim and will determine the role of p65 and IKK in this process. Aim 3 will use in vitro and in vivo models to determine whether regulation of Bim protein stability governs EMT and metastasis in lung cancer cells. Understanding the mechanisms used by inflammatory mediators to stimulate Bim degradation will potentially result in the identification of novel approaches to preventing NSCLC metastasis.

描述（通过应用程序证明）：肺癌会导致最常见的疾病形式更多。 NSCLC肿瘤通常在NSCLC肿瘤中升高NSCLC细胞如何重新分辨出来，我们已经表征了NF-的分子途径？对NSCLC细胞中的Anoikis的尸体蛋白质很重要。直接磷酸化BIM，通过涉及多聚泛素化和蛋白酶体降解的过程来调节其半衰期tgf¿ - 诱导的EMT。 NSCLC转移的速率损失调节BIM蛋白质的转移，并确定该促进性的响应机制是否会解决NSCLC的三个特定目标如果p65的斑点对微管络合物或线粒体诱导的凋亡，则需要刺激BIM降解的最小相互作用。稳定性是否控制着肺癌细胞中的EMT和转移。