Chemoprevention of prostate cancer by resveratrol

白藜芦醇对前列腺癌的化学预防

• 批准号: 6829195
• 负责人: TZE-CHEN HSIEH
• 金额: $ 7.8万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6829195
• 关键词: Escherichia coli; SDS polyacrylamide gel electrophoresis; carcinogenesis; cell proliferation; chemical structure function; chemoprevention; flavonoids; gene expression; hormone related neoplasm /cancer; intermolecular interaction; mass spectrometry; neoplasm /cancer nutrition therapy; neoplastic cell; nutrition related tag; polymerase chain reaction; prostate neoplasms; protein binding; protein purification; protein quantitation /detection; protein structure function; silver impregnation; stilbenes; western blottings

DESCRIPTION (provided by applicant): This proposal aims to identify cellular targets of a grape-skin derived chemical called resveratrol, in regards to its chemopreventive activities against prostate cancer (CAP). The studies described herein expand on our published observations that this dietary polyphenol significantly reduced growth of androgen responsive (AD) and hormone refractory (AI) CaP cells, and profoundly lowered prostate specific antigen (PSA) expression. The slow natural history of CaP and the long latency associated with AD->AI transition present an ideal model for testing whether this specific agent confers chemoprotection in humans. As a requisite to fulfilling this long-term objective, I propose to identify and characterize cellular targets of resveratrol using a novel approach, which I have named resveratrol-capture proteomics. The underpinning of this innovative approach lies in the aromatic stilbene core of resveratrol and its hydrophilic side groups, which I hypothesize to interact with and specifically bind distinct targeting proteins (herein referred to as resveratrol targeting proteins, RTPs). We have devised an innovative, specific strategy to facilitate the identification of RTPs, in the context of CaP progression. The collective goal of this project therefore is to isolate, identify, clone and express RTPs, using cells mimicking different stages of CaP, for comparison with RTPs derived from normal prostate epithelial and stromal cells. Our experimental approach is to couple resveratrol to epoxy-activated agarose beads, forming a resveratrol-capture affinity matrix. We will test its ability to capture cellular targeting proteins. Specificity of capture will be investigated by varying elution conditions (changing salt, pH, displacement ligands etc.). Proteins binding with the strongest affinity to the matrix will be eluted with resveratrol. They will be checked for purity and then analyzed by mass spectrometry, to ascertain whether they comprise of known proteins in the human genome database. Proteins successfully identified by mass spectrometry will be cloned and expressed. These studies should reveal protein candidates with an obligatory or facilitatory function in chemoprevention of CaP by resveratrol, or in the progression of CaP from a treatable AD form to an AI state, currently without curative therapies.

描述（由申请人提供）： 该提案旨在确定其针对前列腺癌（CAP）的化学预防活性（CAP）的葡萄皮衍生化学化学物质的细胞靶标。此处描述的研究扩展了我们已发表的观察结果，即这种饮食多酚显着降低了雄激素反应（AD）和激素难治性（AI）帽细胞的生长，并大大降低了前列腺特异性抗原（PSA）的表达。 CAP的缓慢自然历史和与AD-> AI转变相关的长潜伏期构成了测试该特定药物是否赋予人类化学保护的理想模型。作为实现这一长期目标的必要条件，我建议使用一种新型方法来识别和表征白藜芦醇的细胞靶标，我将其命名为白藜芦醇捕获蛋白质组学。 这种创新方法的基础在于白藜芦醇及其亲水性侧基的芳族Stilbene核心，我假设它们可以与之相互作用并专门结合不同的靶向蛋白质（本文称为白藜芦醇靶向蛋白，RTPS）。我们已经设计了一种创新的，具体的策略，以促进cap进展的背景下的识别RTP。因此，该项目的集体目标是使用模仿CAP不同阶段的细胞隔离，识别，克隆和表达RTP，以与源自正常前列腺上皮细胞和基质细胞的RTP进行比较。我们的实验方法是将白藜芦醇与环氧激活的琼脂糖珠相结合，形成白藜芦醇捕获亲和矩阵。我们将测试其捕获细胞靶向蛋白的能力。捕获的特异性将通过不同的洗脱条件（改变盐，pH，位移配体等）进行研究。蛋白质与基质具有最强亲和力结合，将用白藜芦醇洗脱。将检查它们的纯度，然后通过质谱分析，以确定它们是否包括人类基因组数据库中已知蛋白质。通过质谱成功鉴定的蛋白质将克隆并表达。这些研究应揭示候选蛋白质在白藜芦醇中具有强制性或促进功能的蛋白质，或者在CAP从可处理的AD形式的CAP进展到目前没有治疗疗法的AI状态。