Genistein and Prostate Cancer

金雀异黄素和前列腺癌

基本信息

批准号：
7454366
负责人：
RAJVIR DAHIYA
金额：
$ 27.16万
依托单位：
NORTHERN CALIFORNIA INSTITUTE/RES/EDU
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-08-09 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7454366
关键词：
APC gene Abbreviations Accounting Androgen Antagonists Androgen Receptor Androgens Apoptosis Asians Benign Binding Sites Biochemical Bioinformatics Biological Assay CD44 gene CDKN1A gene California Cancer Cell Growth Cancer Etiology Candidate Disease Gene Cell Adhesion Cell Cycle Cell Cycle Arrest Cell Cycle Regulation Cell Line Cell Proliferation Cells Cessation of life Chemopreventive Agent Class Clinical Trials Consumption Country Cyclin Gene DNA DNA Methylation DNA Methyltransferase DNA Methyltransferase 3B DNA Methyltransferase Inhibitor DNA Modification Methylases DNA Modification Process DNA glycosylase Data Decitabine Deoxycytidine Dependence Development Diet Dose E-Cadherin Enzyme Gene Enzymes Epigenetic Process Epithelial Cells Estrogen Receptors Estrogens Event GSTP1 gene Gene Expression Genes Genistein Genomics Glutathione S-Transferase Goals Growth Histone Acetylation Histone Deacetylase Histone Deacetylase Inhibitor Histones Hormonal Hormones Implant In Vitro Incidence Isoflavones KAI1 gene LNCaP Laboratories Lead Literature Malignant Neoplasms Malignant neoplasm of prostate Mediating Mentors Messenger RNA Methods Methylation Modification Molecular Mus Northern Blotting Nude Mice Numbers Nutritional PC3 cell line PTEN gene Pathologist Pathway interactions Phytoestrogens Plants Play Poison Polymerase Chain Reaction Prevention Property Prostate Proteins Publishing Radiolabeled Receptor Gene Reporting Research Personnel Retinoblastoma Reverse Transcriptase Polymerase Chain Reaction Reverse Transcription Role San Francisco Scientist Second Primary Cancers Site Small Interfering RNA Societies Soy Proteins Standards of Weights and Measures Steroids System TP53 gene Techniques Testing Therapeutic Time Transferase Trichostatin A Tumor Suppressor Genes Tumor Tissue United States Universities Western Blotting Work base bisulfite bone sialoprotein cancer cell cancer diagnosis cancer prevention cancer therapy cell growth chromatin immunoprecipitation dietary supplements feeding gene repression glutathione S-transferase pi histone acetyltransferase human HDAC4 protein in vivo mRNA Expression male men novel novel strategies nutrition oncoprotein p21 p27 Cell Cycle Protein p27 Enzyme Inhibitor prevent programs promoter prostate cancer prevention radiotracer research study response soy transcription factor trichostatin C tumor tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): The main goal of this proposal is to investigate whether genistein inhibits prostate cancer growth through epigenetic pathways. The rationale for this project is that Western men have a 5-6 fold higher incidence of prostate cancer than Asians. One reason for this discrepancy may be the high soy diet (genistein) consumed by Asians. We hypothesize that genistein inhibits prostate cancer growth through epigenetic pathways. Specific Aim # 1. To investigate whether genistein can suppress prostate cancer growth through cell cycle arrest, cyclin genes and apoptosis. Based on the published literature and preliminary data, we lypothesize that genistein will inhibit prostate cancer cell growth in vitro. We will investigate whether genistein alone or in combination with demethylating agent (5-'-aza-2-deoxycytidine (5-AZA-C) and histone deacetylase inhibitor (trichostatin (TSA) can inhibit prostate cancer growth in vitro. A panel of human prostate cell lines will be studies for cell proliferation, cell cycle, cyclin genes and apoptosis. Specific Aim # 2. To investigate the molecular mechanisms of genistein action through epigenetic pathways. Based on our preliminary data, we hypothesize that genistein inhibits prostate cancer growth through modulation of epigenetic events, such as DNA methylation and histone acetylation. To test this hypothesis, we will analyze: 1) Global methylation and enzymatic activity of DNA methyltransferase (DNMT) using radiolabeled S- adenosylmethionine (SAM) assays. 2) Promoter methylation of candidate genes using bisulfite modification of DNA based techniques. 3) Alterations in gene-specific histone acetylation using ChIP assay, and enzymatic activities of histone acetyl transferases (HATs). 4) Histone deacetylase (HDACs) analysis using biochemical assays. 5). Alterations in mRNA expression of genes that encode enzymes participating in DNA methylation and histone acetylation including DNMTs, demethylases, HATs, HDACs using RT-PCR and northern blot assays. 6) To analyze transcription factors which mediate the effects of genistein on prostate cancer. Specific Aim # 3. To investigate whether genistein has anti-tumor effects through reversal of epigenetic pathways in prostate cancer using an in vivo system. Based on the published literature, genistein or demethylating agents have anti-proliferative effects on various cancers. We hypothesize that genistein will inhibit prostate cancer growth through reversal of epigenetic pathways. Studies will be conducted in athymic nude mice implanted with androgen-responsive LNCaP and androgen nonresponsive DuPro prostate cancer cells. Mice will be fed a diet supplemented with genistein, or a combination of genistein with 5-AZA-C or TSA. The following experiments will be conducted: 1) Determine the in vivo anti-tumor effects of genistein and in combination with 5-AZA-C or TSA. 2) To evaluate expression of steroid receptor genes and tumor suppressor genes at the mRNA and protein levels using Northern blot, and Western blotting analysi respectively. 3) To analyze whether genistein treatment can modulate global and candidate gene methylation in tumor tissue using a radiolabeled SAM assay and bisulfite modified genomic sequencing techniques respectively. 4) To analyze, in tumor tissue, gene-specific histone acetylation using ChIP assay and the enzymatic activities of HATs and HDACs using biochemical assays. Accomplishment of these experiments will demonstrate whether epigenetic events are modulated by genistein and provide rationale for a clinical trial of genistein alone or in combination with other epigenetic modulators for the treatment or prevention of prostate cancer.

描述（由申请人提供）：该提案的主要目标是研究染料碱是否通过表观遗传途径抑制前列腺癌的生长。该项目的理由是，西方男性的前列腺癌发病率比亚洲人高5-6倍。这种差异的原因之一可能是亚洲人消耗的高大豆饮食（染料木黄酮）。我们假设染料碱性通过表观遗传途径抑制前列腺癌的生长。具体目的＃1。研究染料木黄酮是否可以通过细胞周期停滞，细胞周期蛋白基因和凋亡来抑制前列腺癌的生长。根据已发表的文献和初步数据，我们认为染料木黄酮会在体外抑制前列腺癌细胞的生长。我们将研究染料木黄酮单独还是与脱甲基化剂（5 -'- AZA-2-脱氧辛丁胺（5-Aza-C）和组蛋白脱乙酰基酶抑制剂（Trichostatin（TSA）可以抑制体内前的癌症生长。前列腺细胞线将是细胞增殖，细胞周期，细胞周期基因和细胞凋亡的研究。表观遗传事件的调节，例如DNA甲基化和组蛋白乙酰化。使用基于DNA的技术的亚硫酸盐修饰的候选基因。 3）使用CHIP测定法和组蛋白乙酰转移酶（HATS）的酶促活性改变基因特异性组蛋白乙酰化。 4）使用生化测定法分析组蛋白脱乙酰基酶（HDACS）。 5）。使用RT-PCR和Northern印迹测定法编码参与DNA甲基化和组蛋白乙酰化的酶的基因的mRNA表达改变，包括DNMT，脱甲基酶，HATS，HAT，HDACS。 6）分析转录因子介导染料木黄酮对前列腺癌的影响。特定目的＃3。研究染料木黄酮是否通过使用体内系统在前列腺癌中的表观遗传途径逆转是否具有抗肿瘤作用。根据已发表的文献，染料木黄酮或脱甲基剂对各种癌症具有抗增殖作用。我们假设染料木黄酮会通过表观遗传途径的逆转来抑制前列腺癌的生长。研究将在植入雄激素反应性LNCAP和雄激素非反应性DUPRO前列腺癌细胞的无性裸鼠中进行研究。小鼠将被喂食，饮食中补充染料木黄酮，或将染料木黄酮与5-aza-C或TSA组合在一起。将进行以下实验：1）确定染料木黄酮的体内抗肿瘤作用，并与5-Aza-C或TSA结合使用。 2）分别使用Northern blot评估类固醇受体基因和肿瘤抑制基因的表达，并分别使用Northern印迹和蛋白质印迹分析。 3）分析染料木黄酮治疗是否可以分别使用放射性标记的SAM分析和亚硫酸盐修饰的基因组测序技术来调节肿瘤组织中的全球和候选基因甲基化。 4）在肿瘤组织中，使用CHIP测定法和使用生化测定法分析帽子和HDAC的酶促活性。这些实验的完成将证明表观遗传事件是通过染料木黄酮调节的，并为单独的染料木黄酮的临床试验提供了理由，还是与其他表观遗传调节剂结合使用，以治疗或预防前列腺癌。