DEVELOPMENT OF CELL LINES

我们建议建立表达大鼠细胞色素P450的人细胞系 CDNA。编码P450IIB1和P450IID1的两个大鼠细胞色素P450 cDNA将 将其导入人淋巴母细胞系AHH-1TK/-。 含有Epstein Barr病毒复制起点的载体将是 用于这一转化体。重组细胞群体将被分离 和特定的细胞色素P450催化的反应将在 重组细胞群体和对照细胞群体。重组细胞系将 也可用于基因突变检测。已知的原诱变剂被激活 通过对具体的P450表格进行测试。这样做的最终目的是 计划是开发一组表达大鼠细胞色素的细胞系 P450。该仪表板可用于分析大鼠外来化合物代谢。 有能力将代谢产物的形成与诱导 在同一细胞系中的遗传毒性。其他应用包括 菌株或性别特异性细胞色素P450的分析及与 人类P450在同一细胞系统中。

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.