DEVELOPMENT OF CELL LINES

我们建议开发表达大鼠细胞色素P450的人细胞系 cDNA。两个大鼠细胞色素P450 cDNA，编码P450 IIB 1和P450 IID 1， 转染入AHH-1 TK+/-人淋巴母细胞系。 含有爱泼斯坦巴尔病毒复制起点的载体将被 在这次转染中使用。 将分离重组细胞群 和特定的细胞色素P450催化的反应将在 重组和对照细胞群体。 重组细胞系将 也可用于基因突变分析。 已知激活的前诱变剂 将测试特定P450形式。 这个项目的最终目标 计划是开发一组表达大鼠细胞色素 P450 该面板可用于分析大鼠外源化合物代谢 具有将代谢物形成与诱导 在同一细胞系中的遗传毒性。 另外的应用包括 分析菌株或性别特异性细胞色素P450，并与 人类P450在同一个细胞系统中。

We propose to develop human cell lines expressing rat cytochrome P450 cDNAs. Two rat cytochrome P450 cDNAs, encoding P450IIB1 and P450IID1, will be transfected into the AHH-1 TK+/- line of human lymphoblastoid cells. Vectors containing the Epstein Barr virus origin of replication will be used in this transfection. Recombinant cell populations will be isolated and specific cytochrome P450 catalyzed reactions will be measured in recombinant and control cell populations. The recombinant cell lines will also be used in gene mutation assays. Promutagens known to be activated by the specific P450 forms will be tested. The eventual goal of this program is to develop a panel of cell lines expressing rat cytochrome P450s. This panel can be used to analyze rat foreign compound metabolism with the ability to relate metabolite formation with the induction of genotoxicity in the same cell line. Additional applications include analysis of strain or sex specific cytochrome P450s and also comparison to human P450s in the same cell system.