DNA methylation in aging, race and prostate cancer

衰老、种族和前列腺癌中的 DNA 甲基化

• 批准号: 7092089
• 负责人: RAJVIR DAHIYA
• 金额: $ 32.22万
• 依托单位: NORTHERN CALIFORNIA INSTITUTE/RES/EDU
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-15 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7092089
• 关键词: African American; CD44 molecule; CpG islands; DNA methylation; Japanese; aging; amidohydrolases; biomarker; cadherins; caucasian American; clinical research; enzyme activity; estrogen receptors; gene expression; glutathione transferase; human middle age (35-64); human old age (65+); human tissue; male; methyltransferase; molecular oncology; neoplasm /cancer genetics; nucleic acid sequence; patient oriented research; prostate neoplasms; racial /ethnic difference; restriction fragment length polymorphism; tumor suppressor genes

The main goal of this proposal is to identify and characterize the epigenetic alterations that are involved in aging and race-related prostate cancer. Specific hypotheses: 1) The DNA methylation status of most frequently deleted genes [ERs, E-cadherin, CD44 and GSTPI (glutathione S-transferase pi)] will be different in aging and race-related prostate cancer. 2) Identification of novel DNA methylated genes will provide us the potential biomarkers of aging and race-related prostate cancer. 3) DNA methyltransferase, DNA demethylase and histone deacetylase activities will correspond to DNA methylation status in aging and race-related prostate cancer. Specific Aim number 1. To test the hypothesis that inactivation of ER genes by DNA methylation can be used as biomarkers for aging and race-related prostate cancer. We hypothesize that ERalpha and ERbeta are differentially expressed in different aging and race related prostate cancer. To test this hypothesis we will investigate the mRNA and protein expression of ER types (alpha and beta) in different stages and grades of prostate cancer. Protein expression of ERs will be analyzed by immunohistochemistry (for localization) and western blotting (for quantitation). RT-PCR (for screening) and northern blotting (for quantitation) will analyze Gene expression. DNA methylation will be analyzed by sodium bisulfite methylation techniques and confirm by direct DNA sequencing. Specific Aim number 2. To test the hypothesis that inactivation of E-cadherin, CD44 and GSTPI genes by DNA methylation can be used as biomarkers for aging and race-related prostate cancer. Based on our preliminary data and prior publications, E-cadherin, CD44 and GSTPI genes are most frequently inactivated by DNA methylation in prostate cancer. We hypothesize that the expression or inactivation of these genes will be different in aging and race- related prostate cancer. To test this hypothesis, we will first determine the protein expression of these genes in prostate cancer using immunohistochemistry (for localization), and western blotting (for quantitation). RNA expression will be analyzed by RT-PCR (for screening) and northern blotting (for quantitation). CpG methylation will be analyzed by sodium bisulfite methylation techniques and confirm by direct DNA sequencing. Specific Aim number 3. To test the hypothesis that identification of new DNA methylated genes by restriction landmark genomic scanning (RLGS) can be used as novel biomarkers for aging and race-related prostate cancer. We hypothesize that identification of novel genes that are inactivated by DNA methylation will be important in understanding the mechanisms of aging and race-related prostate cancer. To test this hypothesis, we will analyze new genes that are methylated in aging and race-related prostate cancer. We will use Restriction landmark genomic scanning for methylation (RLGS) to survey genome wide methylation alterations in aging and race-related prostate cancer and subsequently verify potentially hypermethylated gene loci using sodium bisulfite sequencing technique. Specific Aim number 4. To investigate the mechanisms of DNA methylation in aging and race-related prostate cancer. We hypothesize that DNA methyltransferase, DNA demethylase, histone deacetylase are responsible for regulation of DNA hypermethylation. To test this hypothesis, we will analyze DNA methyltransferase, DNA demethylase and histone deacetylase enzyme activity, mRNA and protein expression in different age and race related prostate cancer. Accomplishment of these experiments will demonstrate whether DNA methylated genes are involved in aging and race-related prostate cancer. This information can be used for better management of prostate cancer using DNA methylated genes as diagnostic or prognostic biomarkers.

该提案的主要目标是识别和表征与衰老和种族相关的前列腺癌有关的表观遗传改变。 具体假设：1）最常缺失的基因[ERs、E-cadherin、CD44和GSTPI（谷胱甘肽S-转移酶pi）]的DNA甲基化状态在衰老和种族相关的前列腺癌中会有所不同。 2）新的DNA甲基化基因的鉴定将为我们提供衰老和种族相关前列腺癌的潜在生物标志物。 3) DNA甲基转移酶、DNA去甲基化酶和组蛋白去乙酰化酶活性将与衰老和种族相关前列腺癌中的DNA甲基化状态相对应。 具体目标 1. 检验以下假设：DNA 甲基化导致的 ER 基因失活可用作衰老和种族相关前列腺癌的生物标志物。我们假设 ERalpha 和 ERbeta 在不同衰老和种族相关的前列腺癌中存在差异表达。为了检验这一假设，我们将研究不同阶段和级别的前列腺癌中 ER 类型（α 和 β）的 mRNA 和蛋白质表达。 ER 的蛋白质表达将通过免疫组织化学（用于定位）和蛋白质印迹（用于定量​​）进行分析。 RT-PCR（用于筛选）和 Northern blotting（用于定量​​）将分析基因表达。 DNA 甲基化将通过亚硫酸氢钠甲基化技术进行分析，并通过直接 DNA 测序进行确认。 具体目标 2. 检验以下假设：DNA 甲基化导致的 E-钙粘蛋白、CD44 和 GSTPI 基因失活可用作衰老和种族相关前列腺癌的生物标志物。 根据我们的初步数据和之前的出版物，在前列腺癌中，E-钙粘蛋白、CD44 和 GSTPI 基因最常因 DNA 甲基化而失活。 我们假设这些基因的表达或失活在衰老和种族相关的前列腺癌中会有所不同。 为了检验这一假设，我们将首先使用免疫组织化学（用于定位）和蛋白质印迹（用于定量​​）确定这些基因在前列腺癌中的蛋白质表达。 RNA 表达将通过 RT-PCR（用于筛选）和 Northern 印迹（用于定量​​）进行分析。 CpG 甲基化将通过亚硫酸氢钠甲基化技术进行分析，并通过直接 DNA 测序进行确认。 具体目标 3. 检验以下假设：通过限制性标志基因组扫描 (RLGS) 鉴定新的 DNA 甲基化基因可用作衰老和种族相关前列腺癌的新型生物标志物。 我们假设，鉴定因 DNA 甲基化而失活的新基因对于理解衰老和种族相关前列腺癌的机制非常重要。 为了检验这一假设，我们将分析在衰老和种族相关的前列腺癌中甲基化的新基因。 我们将使用限制性标志基因组甲基化扫描（RLGS）来调查衰老和种族相关前列腺癌的全基因组甲基化改变，并随后使用亚硫酸氢钠测序技术验证潜在的高甲基化基因位点。 具体目标 4：研究衰老和种族相关前列腺癌中 DNA 甲基化的机制。 我们假设DNA甲基转移酶、DNA脱甲基酶、组蛋白脱乙酰酶负责DNA高甲基化的调节。 为了检验这一假设，我们将分析不同年龄和种族相关前列腺癌中的 DNA 甲基转移酶、DNA 脱甲基酶和组蛋白脱乙酰酶活性、mRNA 和蛋白质表达。这些实验的完成将证明 DNA 甲基化基因是否与衰老和种族相关的前列腺癌有关。 该信息可用于使用 DNA 甲基化基因作为诊断或预后生物标志物更好地管理前列腺癌。

项目成果

DNA mismatch repair gene MLH1 induces apoptosis in prostate cancer cells.

• DOI: 10.18632/oncotarget.2315
• 发表时间: 2014-11-30
• 期刊: Oncotarget
• 作者: Fukuhara S;Chang I;Mitsui Y;Chiyomaru T;Yamamura S;Majid S;Saini S;Hirata H;Deng G;Gill A;Wong DK;Shiina H;Nonomura N;Dahiya R;Tanaka Y
• 通讯作者: Tanaka Y

Restoration of insulin-like growth factor binding protein-related protein 1 has a tumor-suppressive activity through induction of apoptosis in human prostate cancer.

• 发表时间: 2003-11
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: K. Mutaguchi;H. Yasumoto;K. Mita;A. Matsubara;H. Shiina;M. Igawa;R. Dahiya;T. Usui