GENETICS OF PAH METABOLISM: ROLE IN LUNG CANCER

本研究的总体目标是：（a）研究苯并（a）芘 (BP)通过容易获得的人组织和细胞系（例如， 胎盘、淋巴细胞、单核细胞和RPMI-1788细胞）和细胞色素 从这些来源分离的P450（s）;（B）制备cDNA克隆用于 多环芳烃诱导型人类细胞色素P450（s）;（c）研究 人类基因;和（d）使用这些cDNA克隆来确定关系 AHH诱导表型与香烟易感性的关系 吸烟诱发的肺癌 多环芳烃（PAH）诱导的细胞色素P450 mRNA，待用特异性或交叉反应抗体鉴定 针对细胞色素P450，将从胎盘中分离， 吸烟者和从淋巴母细胞样细胞系RPMI-1788细胞中诱导， 苯并（a）芘代谢活性的体外研究。 mRNA将 转录成cDNA，将其克隆到E.杆菌 的积极 克隆将通过杂交和蛋白质翻译来鉴定 技术。 cDNA克隆将被用于研究组织， 人类细胞色素P450基因组;转录和 RPMI-1788细胞中的翻译产物; PAH诱导信息;以及在旨在定义 AHH诱导表型与AHH易感性的关系 吸烟导致的肺癌 这些研究中拟使用的各种方法包括 免疫学技术参与的提高和鉴定 特异性抗体、HPLC等蛋白质和化学分离 技术、细胞培养和重组DNA方法。

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.