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和结合靶蛋白（PI：Erxi Wu博士） 项目摘要 胰腺癌（PC）是一种致命疾病，其5年生存率约为6％ 诊断和耐药性。 PC中癌症干细胞（CSC）的存在被认为是主要的 导致PC治疗耐药性和PC患者继电器的原因。盐霉素，最广泛的 已发现使用的球固醇对CSC具有深远的效率，并克服了多种药物 癌症的抗性。我们的初步数据表明，盐霉素对PC具有强大的细胞毒性 细胞。我们确定了两个盐霉素的潜在结合靶标：转录中间因子1Beta（TIF1¿） 和PC细胞中的核苷（NCL）。但是，PC中盐霉素的作用机理仍然不清楚。 尤其是其直接绑定目标。在这个项目中，我们建议盐霉素对 癌症和CSC可能是由于1）细胞增殖和生存的减少和/或2）诱导 细胞分化。这项拟议的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调节的信号通路及其结合蛋白。我们 假设特定靶蛋白存在于盐霉素反应性细胞中，而盐霉素会引发 它通过其结合靶蛋白的功能。将使用三个特定目标来检验假设。目标1 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。盐霉素与TIF1B和NCL的直接结合将使用 免疫沉淀和免疫结合方法。盐霉素及其潜力之间的相互作用 通过分析其关联和解离轮廓，将进一步确认目标。临床相关性 通过评估两个基因的表达水平与 患者的结果。目标2。确定盐霉素对吉西他滨的TIF1和NCL的作用 抗性PC细胞。在各种情况下，将在体外和体内分析盐霉素的影响 TIF1¿和/或NCL的缺乏或过表达。盐霉素的效率以及与 PC上的吉西他滨将使用转基因小鼠模型确定。目标3。剖析信号 盐霉素，吉西他滨调节的途径，它们的组合和盐霉素的结合蛋白 PC细胞。盐霉素及其靶蛋白对基因在关键途径中表达的影响 PC进程将使用自定义的RT-PCR阵列确定。确定的目标将进一步 使用多种策略进行评估。这项拟议的研究将为PC机理提供新的见解 治疗耐药性和使用盐霉素或与宝石组合作为新型抗PC的理论基础 方案。 ____