Curcumin inhbits BPDE-induced damage by lowering the threshold of p53 activation

姜黄素通过降低 p53 激活阈值抑制 BPDE 引起的损伤

• 批准号: 7846237
• 负责人: Erica Nicole Rogers
• 金额: $ 2.41万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7846237
• 关键词: 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; Affect; Americas; Benzo(a)pyrene; Biological Assay; Cancer Etiology; Carcinogens; Cell Cycle Arrest; Cell Cycle Progression; Cells; Cessation of life; Curcumin; DNA; DNA Adducts; DNA Damage; DNA Repair; DNA repair protein; Dietary Component; Disease; Dose; Drug Metabolic Detoxication; Epoxy Compounds; Exposure to; Genes; Genomics; Glutathione; Glutathione S-Transferase; Lesion; Lung; Malignant neoplasm of lung; Mediating; Mediation; Methods; Monitor; Mutagenesis; Pathway interactions; Proteins; Pyrenes; Regulation; Relative (related person); Role; Route; Southern Blotting; Spices; System; TP53 gene; Tobacco smoke; Tobacco-Associated Carcinogen; Western Blotting; benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; carcinogenesis; mortality; prevent; protein expression; repaired; response; 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; Affect; Americas; Benzo(a)pyrene; Biological Assay; Cancer Etiology; Carcinogens; Cell Cycle Arrest; Cell Cycle Progression; Cells; Cessation of life; Curcumin; DNA; DNA Adducts; DNA Damage; DNA Repair; DNA repair protein; Dietary Component; Disease; Dose; Drug Metabolic Detoxication; Epoxy Compounds; Exposure to; Genes; Genomics; Glutathione; Glutathione S-Transferase; Lesion; Lung; Malignant neoplasm of lung; Mediating; Mediation; Methods; Monitor; Mutagenesis; Pathway interactions; Proteins; Pyrenes; Regulation; Relative (related person); Role; Route; Southern Blotting; Spices; System; TP53 gene; Tobacco smoke; Tobacco-Associated Carcinogen; Western Blotting; benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; carcinogenesis; mortality; prevent; protein expression; repaired; response

DESCRIPTION (provided by applicant): Lung cancer, the leading cause of cancer-related deaths in America, is most commonly caused by long-term exposure to tobacco smoke carcinogens such as benzo[a]pyrene (BaP), which is metabolized into the ultimate metabolite benoz[a]pyrene-7,8-dihyrodiol-9,10-epoxide (BPDE). Despite improvements in the treatment and management of lung cancer, these methods fail to reduce the overall mortality rate of this disease. While dietary components such as curcumin (turmic spice) show promising effects against BPDEinduced carinogenesis, its exact mode of action is not fully understood. The objective of this project is to elucidate whether pretreatment with curcumin inhibits BPDE-induced DMA damage by activating p53. We hypothesize that curcumin may reduce the mutagenic activity of low dose BPDE by lowering the threshold of p53 activation, thereby inducing DNA repair and cell cycle arrest at lower BPDE exposures. In Specific aim 1 the expression of p53 and p53-regulated proteins as well as cell cycle progression will be monitored by Western blot and flow cytometric analysis, respectively, to determine whether curcumin influences cell cycle arrest in response to BPDE exposure. Because global genomic repair (GGR) removes most BPDE-DNA adducts in a p53-dependent manner, Specific aim 2 will use immunoslot blot, [32P]-postlabeling, and UvrABC mediated gene and strand-specific southern blot assays to analyze whether curcumin affects p53 activation and thus the relative extent of BPDE-DNA adducts formed and removed in genomic DNA. In Specific aim 3 Northern and Western blot assays will be used to investigate the role of curcumin in the induction of damage recognition proteins of the GGR pathway (XPC, DDB2), which are p53-dependent. Furthermore it is believed that conjugation of glutathione (GSH) to BPDE by glutathione-S-transferase (GST) interacts with the expression of p53 to inactivate BPDE. In Specific aim 4, Western blot and GST colorimetric assays will be used to monitor the expression of proteins involved in the GSH detoxification system as well as the overall activity of GST, respectively. This aim will provide a better understanding of potential alternative routes that may inactivate BPDE and how curcumin may influence this activity. By investigating the effects of curcumin on p53-mediated responses to DNA damage, a better understanding of the potential role of curcumin in preventing tobacco carcinogen-induced mutagenesis and carcinogenesis will be gained. In addition, the results from this project will benefit the public by providing easy and inexpensive alternative methods in preventing initial and continuing signs of lung cancer.

描述（由申请人提供）：肺癌是美国与癌症相关死亡的主要原因，最常见的是长期暴露于烟草烟雾致癌物（例如苯并[a] pyrene（BAP）），该毒素（BAP）被代谢为最终的代谢物苯甲酸酯[A] pyrene-7,7,7,7,-dihyrodiol-9-10-10-10-110-eppdeide。尽管肺癌的治疗和治疗方法有所改善，但这些方法无法降低该疾病的总死亡率。虽然诸如姜黄素（毛毛香料）之类的饮食成分显示出对BPDESECASINE的富症的有希望的作用，但其确切的作用方式尚未完全了解。该项目的目的是阐明姜黄素预处理是否通过激活p53抑制BPDE诱导的DMA损伤。我们假设姜黄素可以通过降低p53激活的阈值来降低低剂量BPDE的诱变活性，从而在较低的BPDE暴露下诱导DNA修复和细胞周期停滞。在特定的目标1中，将分别通过蛋白质印迹和流式细胞仪分析来监测p53和p53调节的蛋白质以及细胞周期进程的表达，以确定姜黄素是否会影响响应BPDE暴露的细胞周期停滞。因为全球基因组修复（GGR）消除了大多数BPDE-DNA 以p53依赖性的方式加合物，特定的目标2将使用免疫斜率印迹，[32p] - postlabeling和 UVRABC介导的基因和链特异性的南部印迹分析，以分析姜黄素是否影响p53激活，从而在基因组DNA中形成并去除的BPDE-DNA加合物的相对程度。在特定的目标中，将使用北印迹分析和蛋白质印迹分析来研究姜黄素在诱导GGR途径（XPC，DDB2）的损伤识别蛋白中的作用，这些蛋白是p53依赖性的。此外，人们认为通过谷胱甘肽-S-转移酶（GST）将谷胱甘肽（GSH）与BPDE结合与p53的表达相互作用，以使BPDE失活。在特定的目标4中，将使用Western印迹和GST比色测定法分别监测参与GSH解毒系统和GST的整体活性的蛋白质的表达。该目标将更好地理解可能使BPDE失活以及姜黄素如何影响这一活动的潜在替代途径。通过研究姜黄素对p53介导的对DNA损伤的反应的影响，可以更好地理解姜黄素在防止烟草致癌诱导的诱变和致癌作用中的潜在作用。此外，该项目的结果将通过为防止肺癌的初始和持续迹象提供简单且廉价的替代方法来使公众受益。