GENETICS OF PAH METABOLISM: ROLE IN LUNG CANCER

我们调查的总体目标是：(A)研究苯并(A)芘 (BP)通过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 从这些来源分离的P450(S)； 多环芳烃诱导的人细胞色素P450(S)；(C)研究人细胞色素P450的分子生物学 人类基因；及(D)使用这些cdna克隆来确定 AHH诱导性表型与香烟易感性的关系 吸烟导致的肺癌。 多环芳烃(PAH)诱导的细胞色素P450 信使核糖核酸(S)，用特异性或交叉反应抗体鉴定 抗细胞色素P450，将从胎盘中分离出来的 吸烟者和来自淋巴母细胞系RPMI-1788细胞的诱导 体外测定苯并(A)芘的代谢活性。信使核糖核酸将 转录成c DNA，然后克隆到大肠杆菌中。积极的一面 克隆将通过杂交和蛋白质翻译来鉴定 技术高手。这些cDNA克隆将被用于研究 人类细胞色素P450基因组；转录和转录的遗传调节 RPMI-1788细胞中的翻译产物；个体间差异 多环芳烃诱导的信息；在旨在定义 AHH诱导性表型与肺炎易感性的关系 香烟烟雾导致的肺癌。 这些研究中打算使用的各种方法包括 在饲养和鉴定中涉及的免疫学技术 特异性抗体、高效液相等蛋白质和化学分离 技术、细胞培养和重组DNA方法学。

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.