Defining PAH-mediated carcinogenesis in lung: CYP metabolism and B[a]P transport

定义 PAH 介导的肺癌致癌作用：CYP 代谢和 B[a]P 转运

基本信息

批准号：
7502209
负责人：
Stacy Lynn Gelhaus
金额：
$ 4.96万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-30 至 2010-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7502209
关键词：
2&apos -deoxyadenosine ABCB1 gene ABCC1 gene Address Adipose tissue Amines Amino Acids Animals Area Aromatic Polycyclic Hydrocarbons Benz(a)Anthracenes Benzo(a)pyrene Binding Biological Biological Assay CYP1A1 gene CYP3A4 gene CYP3A5 gene Cancer Etiology Carcinogens Cations Cattle Cause of Death Cell Line Cell Nucleus Cells Characteristics Coal CpG Islands Cultured Cells Cytochrome P450 Cytochromes DNA DNA Adduction DNA Adducts DNA Damage Data Deoxyguanosine Development Dimethyl Sulfoxide Dioxins Drug Delivery Systems Drug Metabolic Detoxication Environmental Carcinogens Environmental Pollutants Enzymes Epoxide hydrolase Experimental Models Exposure to Family member Fire - disasters Food Garbage Gases Genomics Glutathione Glutathione S-Transferase Glycol Glycoproteins Guanine Hepatocyte Human Human Milk Hydrolysis In Vitro Individual Individual Differences International Agency for Research on Cancer Itraconazole Kidney Link Liquid Chromatography Literature Liver Lung Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of urinary bladder Mass Spectrum Analysis Meat Mediating Messenger RNA Metabolic Metabolic Activation Metabolic Pathway Metabolism Methodology Methods Modeling Monitor Multidrug Resistance Associated Protein 1 Mus Mutation Names Nature Null Lymphocytes Oils Oxides P-Glycoprotein Pathway interactions Pharmaceutical Preparations Phase Play Point Mutation Promoter Regions Protein Isoforms Protein p53 Proteomics Public Health Pyrenes RNA Interference Reaction Reporting Research Reverse Transcriptase Polymerase Chain Reaction Role Skin Small Interfering RNA Smoke Smoker Stable Isotope Labeling TP53 gene Testing Tobacco Tobacco smoke Toxic effect Transfection Transferase United States United States Dept. of Health and Human Services Western Blotting Wood material Workplace adduct anticancer research base benzanthracene carbonyl reductase (NADPH)carcinogenesis cigarette smoking cigarette smoking cytochrome P450 3A extracellular fluoranthene hepatoma cell inhibitor/antagonist insight lung carcinogenesis multidrug resistance-associated protein 2 novel novel therapeutics prevent protein expression pyrene research study stable isotope tobacco exposure transcription factor

项目摘要

DESCRIPTION (provided by applicant): In this proposed research, benzo[a]pyrene (B[a]P) and it's metabolites, will be used as model compounds to determine the major metabolic pathways of PAHs in lung carcinogenesis. It is generally accepted that cytochrome P4501A1, CYP1A1, is primarily responsible for the metabolism of B[a]P to 7,8-dihydroxy-9,10- epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (B[a]PDE), the ultimate DNA-adduct forming, carcinogenic reactive metabolite; thus connecting CYP1A1 to the toxicological effects of this environmental carcinogen. DNA-adduct formation has been detected at CpG islands located in the promoter region of the tumor suppressor p53. Inactivation of p53 by point mutations is seen in over 40% of all cancers. However, in preliminary research using H358, human bronchoalveolar lung cells, it was found that DNA-adduct levels were higher in non-treated CYP1A1/1B1 null cells compared to H358 cells in which CYP1A1 expression was induced with 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD). This data correlates with literature results of CYP1A1 KO mice stuides in which it was determined that CYP1A1 conferred a protective rather than a toxic effect. This is indicative of the metabolic activity of another CYP in the metabolism of B[a]P. Several other CYP enzymes are capable of catalyzing this metabolism. Preliminary data has also indicated that TCDD may have an effect on the transport of B[a]P-reactive metabolites. It is known that TCDD is able to induce several of the ABCC drug transporter family members; therefore, explaining why less DNA-adduct formation is observed in lung cells pretreated with TCDD . This proposal, which has two aims, will identify the predominate CYP responsible for B[a]P metabolism in the lung and determine the role of B[a]PDE transporters in DNA-adduct formation in lung cells. A stable isotope dilution LC-MRM/MS method, previously developed, will be used for DNA-adduct quantification. DNA-adduct levels will be correlated to mRNA and protein expression determined by RT-PCR and a novel quantitative proteomics method, SI LAC. CYPs including 1A1, 1B1, 1A2, 2B6, 2E1, 2J2, 3A4, and 3A5 will be investigated. DNA-adduct formation will also be monitored in lung cells in which drug transporter expression has been inhibited by siRNA. Continuing research in this area will provide exciting new therapeutic strategies (drug targets) that can be employed to prevent PAH-mediated DNA damage. Relevance to public health: Cigarette smoke is known to cause 90% of lung cancer, but only 10% of smokers develop lung cancer. Examination of alternative CYP metabolism and B[a]PDE transport in lung cells will provide an experimental model to explore whether inter-individual differences in CYP metabolism and transport contribute to these phenomenon.

描述（由申请人提供）：在这项拟议的研究中，苯并[A] pyrene（B [a] P）及其代谢物将用作模型化合物，以确定肺癌中PAH的主要代谢途径。人们普遍认为，细胞色素p4501a1，CYP1A1主要负责B [a] p至7,8-二羟基9,10-9,10-环氧-7,8,9,9,9,10-二氢苯乙烯[a] pyrene [a] pyrene（b [a] pde），Metabimate dna-dna-ddna-ddagogend，因此将CYP1A1与该环境致癌的毒理学作用联系起来。在肿瘤抑制p53的启动子区域的CpG岛上已经检测到DNA-ADDUCT形成。在所有癌症中，超过40％逐点突变p53灭活。 However, in preliminary research using H358, human bronchoalveolar lung cells, it was found that DNA-adduct levels were higher in non-treated CYP1A1/1B1 null cells compared to H358 cells in which CYP1A1 expression was induced with 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD).该数据与CYP1A1 KO小鼠的文献结果相关，其中确定CYP1A1赋予了保护性而不是有毒作用。这表明B [a] p的代谢中另一种CYP的代谢活性。其他几种CYP酶也能够催化这种新陈代谢。初步数据还表明，TCDD可能对B [A] P反应代谢产物的运输有影响。众所周知，TCDD能够诱导几个ABCC毒品转运蛋白家族成员。因此，解释了为什么在用TCDD预处理的肺部细胞中观察到较少的DNA-ADDUCT形成。该提议具有两个目标，将确定负责肺中B [A] P代谢的主要CYP，并确定B [A] PDE转运蛋白在肺细胞中DNA-ADDUCT形成中的作用。先前开发的稳定的同位素稀释LC-MRM/MS方法将用于DNA-ADDUCT定量。 DNA-ADDUCT水平将与通过RT-PCR确定的mRNA和蛋白质表达相关，并将一种新型的定量蛋白质组学方法Si Lac相关。将研究包括1A1、1B1、1A2、2B6、2E1、2J2、3A4和3A5在内的CYP。在肺部抑制药物转运蛋白表达的肺细胞中，还将监测DNA-ADDUCT的形成。在该领域的继续研究将提供令人兴奋的新治疗策略（药物靶标），可用于防止PAH介导的DNA损伤。与公共卫生有关：众所周知，香烟烟雾会导致90％的肺癌，但只有10％的吸烟者患上肺癌。检查肺部细胞中替代CYP代谢和B [A] PDE转运的研究将提供一个实验模型，以探索CYP代谢和运输的个体间差异是否有助于这些现象。