Chemoprevention of prostate cancer by resveratrol

白藜芦醇对前列腺癌的化学预防

• 批准号: 6919976
• 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 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6919976
• 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）： 该提案旨在确定一种名为白藜芦醇的葡萄皮衍生化学物质的细胞靶标，了解其对前列腺癌 (CAP) 的化学预防活性。本文描述的研究扩展了我们已发表的观察结果，即这种膳食多酚显着降低了雄激素反应性 (AD) 和激素难治性 (AI) CaP 细胞的生长，并显着降低了前列腺特异性抗原 (PSA) 的表达。 CaP 缓慢的自然历史以及与 AD->AI 转变相关的长潜伏期为测试这种特定药物是否赋予人类化学保护作用提供了一个理想的模型。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的芳香二苯乙烯核心及其亲水性侧基，我假设它们与不同的靶向蛋白（本文称为白藜芦醇靶向蛋白，RTP）相互作用并特异性结合。我们设计了一种创新的具体策略，以促进在 CaP 进展的背景下识别 RTP。因此，该项目的共同目标是使用模拟 CaP 不同阶段的细胞来分离、鉴定、克隆和表达 RTP，以便与源自正常前列腺上皮和基质细胞的 RTP 进行比较。我们的实验方法是将白藜芦醇与环氧活化的琼脂糖珠偶联，形成白藜芦醇捕获亲和基质。我们将测试其捕获细胞靶向蛋白的能力。将通过改变洗脱条件（改变盐、pH、置换配体等）来研究捕获的特异性。与基质具有最强亲和力结合的蛋白质将用白藜芦醇洗脱。将检查它们的纯度，然后通过质谱分析，以确定它们是否包含人类基因组数据库中的已知蛋白质。通过质谱成功鉴定的蛋白质将被克隆和表达。这些研究应揭示候选蛋白质在白藜芦醇化学预防 CaP 中或在 CaP 从可治疗的 AD 形式发展为目前尚无治疗方法的 AI 状态中具有强制性或促进功能。

项目成果

Identification of glutathione sulfotransferase-pi (GSTP1) as a new resveratrol targeting protein (RTP) and studies of resveratrol-responsive protein changes by resveratrol affinity chromatography.

• 发表时间: 2008
• 期刊: Anticancer research
• 影响因子: 2
• 作者: T. Hsieh;Zhirong Wang;Haiteng Deng;Joseph M. Wu