GENETICS OF PAH METABOLISM: ROLE IN LUNG CANCER

PAH 代谢的遗传学：在肺癌中的作用

• 批准号: 3174715
• 负责人: HIRA L GURTOO
• 金额: $ 10.51万
• 依托单位: ROSWELL PARK CANCER INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-03-01 至 1987-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3174715
• 关键词: Escherichia coli; bacterial RNA; benzopyrenes; carbopolycyclic compound; clone cells; complementary DNA; cytochrome P450; gel electrophoresis; gene expression; genetic regulation; genetic transcription; genetic translation; high performance liquid chromatography; human tissue; lung neoplasms; molecular cloning; molecular oncology; neoplasm /cancer genetics; radiotracer

The overall goals of our investigations are to: (a) study benzo(a)pyrene (BP) metabolism by easily available human tissues and cell lines (e.g. placenta, lymphocytes, monocytes and RPMI-1788 cells) and by cytochrome P450(s) isolated from these sources; (b) prepare cDNA clones for PAH-inducible human cytochrome P450(s); (c) study the molecular biology of the human genes; and (d) employ these cDNA clones to define relationship between AHH inducibility phenotypes and susceptibility to cigarette smoke-induced lung cancer. The polycyclic aromatic hydrocarbon (PAH)-inducible cytochrome P450 mRNA(s), to be identified with specific or cross-reacting antibodies against cytochrome P450, will be isolated from placentas derived from smokers and from the lymphoblastoid cell line RPMI-1788 cells induced in vitro for benzo(a)pyrene metabolizing activity. The mRNA will be transcribed to make cDNA which will be cloned in E. coli. The positive clones will be identified by hybridization and protein translation techiques. The cDNA clones will be used to investigate the organization of the human cytochrome P450 genome; genetic regulation of transcriptional and translational products in RPMI-1788 cells; inter-individual variability in the PAH-inducible message; and in investigations designed to define relationship between AHH inducibiltiy phenotypes and susceptibility to cigarette smoke-induced lung cancer. Various methodologies intended to be used in these studies include immunological techniques involved in the raising and identification of specific antibodies, HPLC and other protein and chemical separation techniques, cell culture and recombinant DNA methodologies.

我们研究的总体目标是： (a) 研究苯并(a)芘 （BP）容易获得的人体组织和细胞系（例如， 胎盘、淋巴细胞、单核细胞和 RPMI-1788 细胞）和细胞色素 从这些来源中分离出的 P450； (b) 制备 cDNA 克隆 PAH 诱导型人细胞色素 P450(s)； (c) 研究分子生物学 人类基因； (d) 使用这些 cDNA 克隆来定义关系 AHH 诱导表型与香烟易感性之间的关系 烟雾诱发的肺癌。 多环芳烃 (PAH) 诱导细胞色素 P450 mRNA，用特异性或交叉反应抗体进行识别 抗细胞色素 P450，将从胎盘中分离出来 吸烟者和来自淋巴母细胞系 RPMI-1788 细胞诱导的 体外苯并(a)芘代谢活性。 mRNA 将是 转录产生 cDNA，并将其克隆到大肠杆菌中。 积极的一面 将通过杂交和蛋白质翻译来鉴定克隆 技术。 cDNA 克隆将用于研究 人类细胞色素P450基因组；转录的遗传调控 RPMI-1788细胞中的翻译产物；个体间差异 PAH 诱导信息；并在旨在定义的调查中 AHH诱导表型与易感性之间的关系 香烟烟雾诱发肺癌。 这些研究中使用的各种方法包括 免疫学技术涉及培养和鉴定 特异性抗体、HPLC等蛋白质和化学品分离 技术、细胞培养和重组 DNA 方法。