Effects of salinomycin and binding target proteins in pancreatic cancer

盐霉素和结合靶蛋白对胰腺癌的影响

基本信息

批准号：
8813062
负责人：
Erxi Wu
金额：
$ 23.27万
依托单位：
NORTH DAKOTA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-01 至 2021-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8813062
关键词：
Antineoplastic Agents Binding Binding Proteins Cancer Center Cancer Intervention Cancer Patient Cancer cell line Cell Differentiation Induction Cell Proliferation Cell Survival Cells Data Diagnosis Diagnostic Disease Dissociation Drug Design Gene Expression Genes Genetic Transcription Goals Human Immune Immunoprecipitation In Vitro Investigation Knowledge Malignant Neoplasms Malignant neoplasm of pancreas Molecular Mechanisms of Action Multi-Drug Resistance Mus Pathway interactions Patient-Focused Outcomes Patients Pharmaceutical Preparations Pilot Projects Proteins Public Health Publishing Regimen Relapse Reporting Research Research Project Grants Resistance Reverse Transcriptase Polymerase Chain Reaction Role Signal Pathway Signal Transduction Specimen Structural Models Survival Rate Testing Therapeutic Transgenic Mice Xenograft procedure base cancer cell cancer stem cell cancer therapy chemotherapy clinically relevant cytotoxicity gemcitabine in vitro Assay in vivo insight knock-down mouse model notch protein novel novel anticancer drug novel therapeutics nucleolin overexpression pancreatic cancer cells programs salinomycin therapeutic target transcriptional intermediary factor 1 tumor tumor progression

项目摘要

_________________________________________________________________________________________________________________________ Project-2. Salinomycin's Effects and Binding Target Proteins in Pancreatic Cancer (PI: Dr. Erxi Wu) Project Summary Pancreatic Cancer (PC) is a deadly disease and its 5-year survival rate is approximately 6% due to late diagnoses and therapy resistance. The existence of cancer stem cells (CSCs) in PC is considered as a major cause for PC therapy resistance and PC patients' relapse from therapy. Salinomycin, one of the most widely used coccidiostats, has been found to possess profound efficacy towards CSCs and to overcome multiple drug resistance in cancers. Our preliminary data showed that salinomycin possesses strong cytotoxicity against PC cells. We identified two salinomycin's potential binding targets: transcription intermediary factor-1beta (TIF1¿) and nucleolin (NCL) in PC cells. However, the action mechanism of salinomycin in PC still remains unclear; especially its direct binding targets. In this project, we propose that the inhibitory effects of salinomycin on cancer and CSCs could be due to 1) the reduction of cell proliferation and survival and/or, 2) the induction of cell differentiation. The goal of this proposed study is to determine salinomycin's binding target proteins and their functions in PC as well as the signaling pathways regulated by salinomycin and its binding proteins. We hypothesize that specific target proteins exist in the salinomycin responsive cells and that salinomycin initiates its function via its binding target proteins. Three specific aims will be used to test the hypothesis. Aim 1. To determine the binding target proteins of salinomycin in PC and PC-CSC as well as their clinical relevance using pathological specimens. The direct binding of salinomycin to TIF1b and NCL will be determined using immunoprecipitation and immune-binding approaches. The interaction between salinomycin and its potential targets will be further confirmed by analyzing their association and dissociation profiles. The clinical relevance of TIF1b and NCL will be examined by assessing the correlation of the expression levels of both genes with patients' outcomes. Aim 2. To determine the roles of TIF1¿ and NCL for salinomycin against Gemcitabine resistant PC cells. The effects of salinomycin will be analyzed in vitro and in vivo at various conditions including lack or overexpression of TIF1¿ and/or NCL. The efficacies of salinomycin and the combination with Gemcitabine on PC will be determined using a transgenic mouse model. Aim 3. To dissect the signaling pathways regulated by salinomycin, gemcitabine, their combination, and the binding proteins of salinomycin in PC cells. The effects of salinomycin and its target proteins on the expression of genes in the key pathways in PC progression will be determined using customized RT-PCR array. The identified targets will be further evaluated using multiple strategies. This proposed study will provide new insight into the mechanism of PC therapy resistance and a theoretical basis for the use of salinomycin or combination with Gem as novel anti-PC regimens. _________________________________________________________________________________________________________________________

_________________________________________________________________________________________________________________________________ 项目-2.盐霉素在胰腺癌中的作用和结合靶蛋白（PI：吴二喜博士）项目概要胰腺癌（PC）是一种致命的疾病，由于晚期癌症，其 5 年生存率约为 6% PC 中癌症干细胞 (CSC) 的存在被认为是影响诊断和治疗耐药性的主要原因。盐霉素是 PC 治疗耐药和 PC 患者复发的原因，盐霉素是最广泛使用的药物之一。使用球虫抑制剂，已被发现对 CSC 具有深远的功效并克服多种药物我们的初步数据表明盐霉素对 PC 具有很强的细胞毒性。我们确定了盐霉素的两个潜在结合靶标：转录中间因子-1β (TIF1¿) 然而，盐霉素在PC细胞中的作用机制仍不清楚。在这个项目中，我们提出盐霉素对特别是其直接结合靶点的抑制作用。癌症和癌症干细胞可能是由于 1) 细胞增殖和存活减少和/或 2) 诱导这项研究的目的是确定盐霉素的结合靶蛋白和它们在 PC 中的功能以及盐霉素及其结合蛋白调节的信号通路。特定靶蛋白存在于盐霉素反应细胞中并且盐霉素启动将使用三个特定目标来检验该假设。确定 PC 和 PC-CSC 中盐霉素的结合靶蛋白及其临床相关性使用病理标本确定盐霉素与 TIF1b 和 NCL 的直接结合。免疫沉淀和免疫结合方法之间的相互作用及其潜力。通过分析其关联和解离概况，将进一步确认目标。 TIF1b 和 NCL 的相关性将通过评估这两个基因的表达水平与目标 2. 确定 TIF1 的作用。盐霉素对抗吉西他滨的 NCL 将在体外和体内不同条件下分析盐霉素的作用。 TIF1 缺乏或过度表达¿和/或 NCL 盐霉素及其组合的功效。吉西他滨对 PC 的影响将使用转基因小鼠模型进行测定。目标 3：剖析信号传导。盐霉素、吉西他滨、它们的组合以及盐霉素结合蛋白调节的途径 PC细胞。盐霉素及其靶蛋白对关键通路中基因表达的影响。 PC 进展将使用定制的 RT-PCR 阵列进行进一步确定。这项拟议的研究将为 PC 的机制提供新的见解。盐霉素或与Gem联合作为新型抗PC药物的耐药性及理论依据治疗方案。 _________________________________________________________________________________________________________________________________