Crespi C L, Langenbach R, Penman B W
Gentest Corporation, Woburn, MA 01801.
Prog Clin Biol Res. 1990;340B:97-106.
We have isolated a human cytochrome P450IIE1 cDNA. This cDNA was transfected to AHH-1 human lymphoblasts and recombinant cell lines were isolated. Two means of selection for the presence of the vector were used, resistance to histidinol and resistance to hygromycin. Histidinol selection was found to give higher vector copy number than hygromycin B selection. AHH-1 cells expressing the P450IIE1 cDNA were sensitive to the cytotoxic and mutagenic effects of exposure to DMN at concentrations of 10 to 30 ng/ml for 28 hours. This system is more sensitive to DMN than systems using hepatocyte mediated activation.
我们分离出了一种人类细胞色素P450IIE1 cDNA。将该cDNA转染至AHH-1人淋巴母细胞中,并分离出重组细胞系。使用了两种选择载体存在的方法,即对组氨醇的抗性和对潮霉素的抗性。发现组氨醇选择比潮霉素B选择能产生更高的载体拷贝数。表达P450IIE1 cDNA的AHH-1细胞对在10至30 ng/ml浓度下暴露于DMN 28小时的细胞毒性和诱变作用敏感。该系统比使用肝细胞介导激活的系统对DMN更敏感。