Mechanistic Studies on Lung Cancer Prevention by Tea Polyphenols

茶多酚预防肺癌的机理研究

基本信息

批准号：
7534818
负责人：
CHUNG S. YANG
金额：
$ 31.95万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-12-01 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7534818
关键词：
8-hydroxy-2&apos -deoxyguanosine A/J Mouse Abbreviations Adenocarcinoma Affinity Affinity Chromatography Allografting Animal Model Apoptosis Binding Binding Proteins Biological Assay Blood Bone Morphogenetic Proteins Butanones Cancer Prevention Trial Cancer cell line Collaborations Consumption Dose EGF gene Epidermal Growth Factor Epigallocatechin Gallate Event FK506 binding protein 5 Future Goals Green tea Growth H1299 Human Immunohistochemistry In Situ Nick-End Labeling In Vitro Inhibition of Apoptosis Investigation Label Lead Lung Neoplasms MAPK8 gene Malignant Neoplasms Malignant neoplasm of lung Mass Spectrum Analysis Matrix Metalloproteinases Mediating Minnesota Modeling Molecular Molecular Target Mus Oxidative Stress Phosphotransferases Polyphenon E Premalignant Preventive Proliferating Cell Nuclear Antigen Propidium Diiodide Proteins Proteomics Proto-Oncogene Proteins c-akt Role Signal Transduction Signal Transduction Pathway Smoker Structure of parenchyma of lung Superoxide Dismutase System Tea Testing Thiobarbituric Acid Reactive Substances Transforming Growth Factors Universities Vascular Endothelial Growth Factors Western Blotting Xenograft procedure adenoma angiogenesis base cancer cell cancer prevention carcinogenesis cellular engineering design gain of function gallocatechol human H2AX protein in vivo insulin receptor substrate 1 protein lung cancer prevention lung carcinogenesis polyphenol research study response tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): Our long-term goal is to understand the cancer preventive activities of tea constituents and their applications to human cancer prevention. The cancer preventive activity of tea polyphenols has been demonstrated in animal models, but the mechanisms of action are not clearly understood. This project aims to elucidate the mechanisms of lung cancer prevention by (-)-epigallocatechin-3-gallate (EGCG), the major green tea polyphenol. Our goal is to establish an integrated in vitro and in vivo experimental system to test our central hypothesis that EGCG inhibits lung carcinogenesis by binding to specific target molecules which are vital to carcinogenesis and this binding triggers alterations in signal transduction pathways that lead to growth inhibition and apoptosis of premalignant and malignant cells. In order to effectively test our hypothesis, we plan to use both in vitro and in vivo experimental systems. The specific aims are as follows: 1. Establish and compare in vivo (xenograft & allograft) and in vitro experimental systems with lung cancer cell lines and study the mechanisms of action of EGCG. Lung cancer cell lines H1299 (human) and CL13 and CL30 (mouse) will be studied in culture and in xenografts/allografts. The effects of EGCG treatment on tumor growth, apoptosis, angiogenesis, and related molecular changes (e.g., ERK1/2, AKT, ASK1, JNK, VEGF, and oxidative stress parameters) will be compared in vivo vs. in vitro. 2. Identify direct molecular targets of EGCG by affinity-proteomics and functional studies. To identify direct molecular targets for EGCG, we will use affinity chromatography and mass spectrometry in collaboration with Dr. Zigang Dong at the University of Minnesota. The functional roles of the putative EGCG target proteins in mediating the action of EGCG will be studied with loss or gain of function experiments in engineered cells and in a xenograft/allograft model established in Aim 1. 3. Elucidate the mechanisms of cancer prevention by EGCG in a 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung carcinogenesis model in A/J mice. Adenoma-bearing mice will receive long- or short-term treatment with EGCG. The cellular and molecular changes related to the inhibition of tumor progression will be investigated using immunohistochemistry, Western blots, and other approaches. We will test specific hypotheses developed based on results from Aims 1 and 2 concerning putative EGCG molecular targets and related molecular events. A better understanding of the mechanisms of the cancer preventive action of EGCG and the dose-response relationship will help us to design future lung cancer prevention trials in humans, for example, in ex-smokers. Project Narrative: Green tea has been shown to have cancer preventive activity in animal models. This project aims to study the lung cancer preventive mechanisms of the major green tea polyphenol, (-)-epigallocatechin-3-gallate (EGCG). The results will help us to design future lung cancer prevention trials in humans and to assess the effects of tea consumption on human lung cancer.

描述（由申请人提供）：我们的长期目标是了解茶成分的癌症预防活性及其在人类癌症预防中的应用。茶多酚的癌症预防活性已在动物模型中得到证实，但其作用机制尚不清楚。该项目旨在阐明主要绿茶多酚 (-)-表没食子儿茶素-3-没食子酸酯 (EGCG) 预防肺癌的机制。我们的目标是建立一个集成的体外和体内实验系统来测试我们的中心假设，即EGCG通过与对致癌作用至关重要的特定靶分子结合来抑制肺癌发生，并且这种结合触发信号转导途径的改变，从而导致生长抑制和癌前细胞和恶性细胞的凋亡。为了有效地检验我们的假设，我们计划使用体外和体内实验系统。具体目标如下： 1.建立并比较肺癌细胞系的体内（异种移植和同种异体移植）和体外实验系统，研究EGCG的作用机制。肺癌细胞系 H1299（人）以及 CL13 和 CL30（小鼠）将在培养物和异种移植物/同种异体移植物中进行研究。将在体内与体外比较 EGCG 治疗对肿瘤生长、细胞凋亡、血管生成和相关分子变化（例如 ERK1/2、AKT、ASK1、JNK、VEGF 和氧化应激参数）的影响。 2. 通过亲和蛋白质组学和功能研究鉴定 EGCG 的直接分子靶标。为了确定 EGCG 的直接分子靶标，我们将与明尼苏达大学的 Zigang Dong 博士合作使用亲和色谱法和质谱法。推定的 EGCG 靶蛋白在介导 EGCG 作用中的功能作用将通过工程细胞和目标 1 中建立的异种移植/同种异体移植模型中的功能丧失或获得实验来研究。 3. 阐明 EGCG 预防癌症的机制在 4-(甲基亚硝基氨基)-1-(3-吡啶基)-1-丁酮 (NNK) 诱导的 A/J 小鼠肺癌模型中。患有腺瘤的小鼠将接受 EGCG 的长期或短期治疗。将使用免疫组织化学、蛋白质印迹和其他方法研究与抑制肿瘤进展相关的细胞和分子变化。我们将测试根据目标 1 和 2 的结果提出的具体假设，涉及推定的 EGCG 分子靶标和相关分子事件。更好地了解 EGCG 的癌症预防作用机制和剂量反应关系将有助于我们设计未来的人类肺癌预防试验，例如在戒烟者中。项目叙述：绿茶在动物模型中已被证明具有预防癌症的活性。该项目旨在研究主要绿茶多酚的肺癌预防机制， (-)-表没食子儿茶素-3-没食子酸酯 (EGCG)。研究结果将帮助我们设计未来的人类肺癌预防试验，并评估饮茶对人类肺癌的影响。