Stable expression of human cytochrome CYP2B6 and CYP1A1 in Chinese hamster CHL cells: their use in micronucleus assays

Abstract

With specific designed primers, CYP2B6 and CYP1A1 cDNA were generated by reverse transcription-polymerase chain reaction(RT-PCR) technique performed on total RNAs isolated from human liver and 3-methylcholanthrene(3-MC) induced human amnion FL cells. Cell lines (CHL-2B6 and CHL-1A1) capable of expressing human cytochrome P450(CYP)2B6 and 1A1 were established after transfection of corresponding eukaryotic recombinant expression plasmid with human CYP2B6 and 1A1 cDNA inserts respectively. These cell lines stably expressed the mRNAs and the enzymatic activities corresponding to CYP2B6 and CYP1A1, respectively. Compared with Chinese hamster 1ung(CHL) cells, the micronucleus frequency in CHL-2B6 cells is markedly increased when exposed to nitrosamines, aflatoxin B1(AFB1) and cyclophosphamide (CPA). This is also in CHL-1A1 cells, when exposed to carcinogenic polycyclic aromatic hydrocarbons.