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

• 批准号: 9230404
• 负责人: Erxi Wu
• 金额: $ 21.27万
• 依托单位: NORTH DAKOTA STATE UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9230404
• 关键词: Antineoplastic Agents; Binding; Binding Proteins; Cancer Intervention; Cancer Patient; Cell Differentiation Induction; Cell Line; Cell Proliferation; Cell Survival; Cells; Custom; Data; Diagnosis; Diagnostic; Disease; Dissociation; Drug Design; Drug resistance; Gene Expression; Genes; Genetic Transcription; Goals; Immune; Immunoprecipitation; In Vitro; Investigation; Knowledge; Malignant Neoplasms; Malignant neoplasm of pancreas; Molecular Mechanisms of Action; Multi-Drug Resistance; Pathologic; Pathway interactions; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Pilot Projects; Proteins; Public Health; Publishing; Regimen; Reporting; Research; Research Project Grants; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Pathway; Specimen; Structural Models; Survival Rate; Testing; Therapeutic; Transgenic Mice; Treatment Efficacy; WNT Signaling Pathway; cancer cell; cancer stem cell; cancer therapy; cancer type; chemotherapy; clinically relevant; cytotoxicity; gemcitabine; human stem cells; in vitro Assay; in vivo; insight; knock-down; mouse model; notch protein; novel; novel anticancer drug; novel therapeutics; nucleolin; overexpression; pancreatic cancer cells; programs; relapse patients; salinomycin; therapeutic target; tumor; tumor progression

_________________________________________________________________________________________________________________________ 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及结合靶蛋白（主要研究者：吴二喜博士） 项目摘要 胰腺癌（PC）是一种致命的疾病，由于晚期胰腺癌，其5年生存率约为6%。 诊断和治疗抗性。肿瘤干细胞（cancer stem cells，CSCs）的存在被认为是肿瘤发生的主要原因。 PC治疗抵抗和PC患者治疗后复发的原因。盐霉素，最广泛的 已经发现，使用的抗球虫药对CSC具有深刻的功效，并且克服了多种药物的抑制作用。 抵抗癌症。初步结果表明盐霉素对PC有较强的细胞毒作用 细胞我们确定了盐霉素的两个潜在结合靶点：转录中介因子1 β（TIF 1） 和核仁素（NCL）。然而，盐霉素在PC中的作用机制尚不清楚; 尤其是其直接结合靶点。在这个项目中，我们提出盐霉素的抑制作用， 癌症和CSCs之间的相互作用可能是由于1）细胞增殖和存活的减少和/或，2） 细胞分化这项研究的目的是确定盐霉素的结合靶蛋白， 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。我们 假设盐霉素应答细胞中存在特异性靶蛋白， 通过结合靶蛋白发挥作用。三个具体目标将被用来检验假设。目标1。到 确定盐霉素在PC和PC-CSC中的结合靶蛋白及其临床意义 使用病理标本。盐霉素与TIF 1b和NCL的直接结合将使用 免疫沉淀和免疫结合方法。盐霉素的相互作用及其潜力 将通过分析它们的结合和解离曲线来进一步确认靶标。临床相关性 将通过评估两种基因的表达水平与TIF 1b和NCL的相关性来检查TIF 1b和NCL的表达水平。 患者的结果。目标2.确定TIF 1 <$和NCL对盐霉素对抗吉西他滨的作用 耐药PC细胞。盐霉素的作用将在体外和体内不同条件下进行分析，包括 缺乏或过度表达TIF 1和/或NCL。盐霉素及其联合应用的疗效 将使用转基因小鼠模型测定PC上的吉西他滨。目标3.为了分析信号 由盐霉素、吉西他滨、它们的组合以及盐霉素在大肠杆菌中的结合蛋白调节的途径。 PC细胞。盐霉素及其靶蛋白对大肠杆菌关键途径基因表达的影响 将使用定制的RT-PCR阵列确定PC进展。确定的目标将进一步 使用多种策略进行评估。这项研究将为PC的机制提供新的见解 为盐霉素或与Gem联合应用作为新型抗PC药物提供了理论依据 养生法 _________________________________________________________________________________________________________________________