Epigenetic changes in a PhIP-induced prostate cancer model in hCYP1A mice

hCYP1A 小鼠 PhIP 诱导的前列腺癌模型的表观遗传变化

• 批准号: 8469733
• 负责人: Jayson Xiao-Chen Chen
• 金额: $ 3.4万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8469733
• 关键词: 2-Amino-1-Methyl-6-Phenylimidazo[4,5-b]pyridine; ABCB1 gene; Aberrant DNA Methylation; Age; Atrophic; CDKN2A gene; Cancer Etiology; Carcinogens; Carcinoma; Cessation of life; Cytochrome P-450 CYP1A2; DNA; DNA Adduction; DNA Methylation; DNA Methyltransferase; DNA Modification Methylases; Data; Developed Countries; Development; Diagnosis; Diet; Disease; Dissection; E-Cadherin; Enzymes; Epigenetic Process; Epithelial; Event; Exposure to; Fishes; Future; GSTP1 gene; Gene Expression Regulation; Gene Mutation; Genes; Genomics; Hematoxylin and Eosin Staining Method; Heterocyclic Amines; High temperature of physical object; Human; Hypermethylation; Inflammation; Laboratories; Lasers; Lateral; Lead; Literature; MAP Kinase Gene; MGMT gene; Maintenance; Malignant Neoplasms; Malignant neoplasm of prostate; Manuscripts; Meat; Mediating; Methods; Methylation; Methyltransferase; Modeling; Molecular; Mus; Mutation; Normal Cell; Nucleotides; Oncogenic; Oxidative Stress; Pathogenesis; Pathway interactions; Play; Prostate; Prostatic Intraepithelial Neoplasias; Public Health; Race; Rattus; Research; Reverse Transcriptase Polymerase Chain Reaction; Risk Factors; Role; Signal Pathway; Signal Transduction; Staging; Techniques; Testing; Time; Transcription Factor AP-1; Tumor Suppressor Genes; United States; Up-Regulation; Western Blotting; base; cancer risk; carcinogenesis; cooking; histone modification; in vivo; inhibitor/antagonist; insight; men; novel; promoter; prostate cancer model; prostate cancer prevention; prostate carcinogenesis; research study

DESCRIPTION (provided by applicant): Prostate cancer is one of the most frequently diagnosed malignancies in developed countries and a leading cause of cancer-related death among men in the United States. Prostate cancer involves numerous risk factors, particularly age, race, and diet. Exposure to PhIP, a dietary carcinogen generated from high temperature cooking of meat and fish, has been associated with prostate cancer risk. In this study, we will investigate carcinogenesis of a PhIP-induced prostate cancer model in CYP1A-humanized (hCYP1A). The objectives are to determine the induction mechanism of DNMT1, a major methyltransferase enzyme involved in DNA methylation, and identify the promoter DNA hypermethylation of genes caused by the upregulated level of DNMT1. Based on previous studies and preliminary data, the hypotheses are that DNMT1 is induced by the Ras/MAPK/AP-1 and/or PI3K/Akt signaling pathway and that the upregulation of DNMT1 causes hypermethylation and inactivation of key regulatory and tumor suppressor genes in the prostate of the humanized mice. To test these hypotheses, three specific aims are proposed: (1) elucidate the mechanism of DNMT1 induction in hCYP1A mice at 1, 3, 7, and 14 days after PhIP treatment; (2) identify promoter DNA hypermethylation of key regulatory and tumor suppressor genes caused the elevated level of DNMT1 using methylation PCR array; and (3) investigate the relationship between DNMT1 upregulation in the early time- points (1 & 3 days) and the late stages (30 & 40 weeks) of PhIP-induced carcinogenesis. The proposed experiments will employ standard histological (H&E), immunohistochemical (IHC), and molecular techniques (Western Blotting & quantitative RT-PCR) as well as more specialized methods (Laser Capture Micro- dissection, Methylation Specific PCR, & DNA methylation PCR array). The study will reveal the induction mechanism of DNMT1 and the epigenetic changes caused by the PhIP carcinogen. Determining the molecular mechanism and alterations in the PhIP-treated hCYP1A mice will enhance our understanding of prostate carcinogenesis and cancer development.

描述（由申请人提供）：前列腺癌是发达国家最常诊断的恶性肿瘤之一，也是美国男性癌症相关死亡的主要原因。前列腺癌涉及许多风险因素，特别是年龄，种族和饮食。暴露于PhIP，一种高温烹饪肉类和鱼类产生的膳食致癌物，与前列腺癌风险有关。在这项研究中，我们将研究PhIP诱导的前列腺癌模型中的CYP 1A-人源化（hCYP 1A）的致癌作用。目的是确定DNMT 1的诱导机制，DNMT 1是参与DNA甲基化的主要甲基转移酶，并鉴定由DNMT 1水平上调引起的基因启动子DNA超甲基化。基于先前的研究和初步数据，假设DNMT 1由Ras/MAPK/AP-1和/或PI 3 K/Akt信号通路诱导，并且DNMT 1的上调导致人源化小鼠前列腺中关键调控和肿瘤抑制基因的超甲基化和失活。为了验证这些假设，我们提出了三个具体的目标：（1）阐明PhIP处理后1、3、7和14天hCYP 1A小鼠DNMT 1诱导的机制：（2）使用甲基化PCR阵列鉴定引起DNMT 1水平升高的关键调控和抑癌基因启动子DNA超甲基化;（3）研究DNMT 1在PhIP诱导的肿瘤发生的早期（1 & 3天）和晚期（30 & 40周）上调之间的关系。拟议的实验将采用标准组织学（H&E）、免疫组织化学（IHC）和分子技术（Western Blotting和定量RT-PCR）以及更专业的方法（激光捕获显微切割、甲基化特异性PCR和DNA甲基化PCR阵列）。该研究将揭示DNMT 1的诱导机制和PhIP致癌物引起的表观遗传变化。确定PhIP处理的hCYP 1A小鼠的分子机制和改变将增强我们对前列腺癌发生和癌症发展的理解。