alexa Genotoxicity test system based on p53R2 gene expression in human cells: examination with 80 chemicals.


Journal of Biosensors & Bioelectronics

Author(s): Ohno K, TanakaAzuma Y, Yoneda Y, Yamada T

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Abstract p53R2, which encodes a subunit of ribonucleotide reductase, is activated by DNA damage induced by gamma-ray and ultraviolet irradiation, and also by genotoxic chemicals such as adriamycin. For the purpose of constructing an easy-operating genotoxicity test system using human cell lines, we developed a p53R2-dependent luciferase reporter gene assay, and demonstrated dose-dependent luminescence caused by adriamycin in two human cell lines that express wild-type p53, MCF-7 and HepG2. The performance of this assay system was evaluated with 80 chemicals including those known in the Ames test as genotoxic or non-genotoxic. When the luciferase activity of cells treated with the test sample was over 200\% to that of control cells in a dose-dependent increasing manner, the sample was judged positive as a genotoxic chemical. Forty of 43 Ames-positive chemicals induced luciferase activity in this assay system. Eight Ames-negative chemicals also induced luciferase activity. These eight chemicals are genotoxic in other in vitro test systems using mammalian cells. It is suggested that this assay system can be applied to rapid screening of chemicals for potential human genotoxicity. This article was published in Mutat Res and referenced in Journal of Biosensors & Bioelectronics

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