HYDROCARBON CARCINOGENESIS IN MOUSE AND HUMAN CELLS

这项研究的两个基本目标是：1)生化 谷胱甘肽S转移酶调节功能的表征 亲电性致癌物诱导的细胞毒性和转化事件 代谢物(例如，苯并(A)芘抗7，8-二氢二醇9，10-环氧化物)和2) 确定允许的致癌物诱导的遗传事件池 用于10T1/2细胞转化。我们将使用集成生化 技术(亲和层析和等电点层析)和分子 技术(重组质粒，DNA转染)来表征 苯并(A)芘可转化的10T1/2小鼠细胞中的谷胱甘肽转移酶 稳定整合外源基因增强其解毒功能 将特异性GSH-转移酶同工酶复制到10T1/2细胞中。在我们的 第二个目标，我们将使用细胞技术(细胞转化)和 分子技术(癌基因-质粒法、杂交法 分析)以表征10T1/2细胞的形态和致瘤性 故意转化为已知的异常癌基因或转化为 致癌物质。我们将使用DNA转染、限制性内切酶 分析和核酸杂交以确定一系列遗传事件 它们与10T1/2细胞转化有关。 我们的目标是整合致癌物质的生物化学观察 代谢及其谷胱甘肽转移酶对分子和遗传学的调控 致癌物与DNA在10T1/2细胞中相互作用结果的观察 以加深对肿瘤转化的理解。

The two basic objectives of this research are to: 1) biochemically characterize the function of glutathione (GSH) S-transferase in regulating the cytotoxic and transforming events induced by electrophilic carcinogen metabolites (e.g. benzo(a)pyrene anti 7,8-dihydrodiol 9,10-epoxide), and 2) identify a pool of carcinogen-induced genetic events which are permissive for 10T1/2 cell transformation. We will use integrated biochemical techniques (affinity and isoelectric point chromatography) and molecular techniques (plasmid reconstruction, DNA transfection) to characterize GSH-transferase in benzo(a)pyrene-transformable 10T1/2 mouse cells and to augment its detoxifying function by stably integrating additional cDNA copies of specific GSH-transferase isozymes into 10T1/2 cells. In our second objective, we will use cellular techniques (cell transformation) and molecular techniques (oncogene-plasmid transfection, hybridization analysis) to characterize the morphology and tumorigenicity of 10T1/2 cells purposely transformed with known aberrant oncogenes or transformed with carcinogens. We will use DNA transfection, restriction endonuclease analysis and nucleic acid hybridization to define a pool of genetic events which are linked to 10T1/2 cell transformation. Our aim here is to integrate biochemical observations of carcinogen metabolism and its regulation by GSH-transferase, to molecular and genetic observations of the results of carcinogen-DNA interaction in 10T1/2 cells, in order to develop an understanding of neoplastic transformation.