The cytotoxic and mutagenic effects of 5-bromodeoxyuridine-plus-black-light treatment in cultured mammalian cells

The cytotoxic and mutagenic effects of the incorporation of 5-bromodeoxyuridine (BrdU)_followed by exposure to black light were investigated with Chinese hamster ovary (CHO) cells in cell culture. Mutation induction at the hypoxanthine-guanine phosphoribosyl transferase (hgprt) locus was determined by selection for 6-thioguanine resistant (TG^r) mutants (CHO/HGPRT system). BrdU alone has been shown to be mutagenic only at concentrations of 50 μM or greater. This study was performed in an effort to determine whether BrdU is actually incorporated into the hgprt gene when lower, nonmutagenic concentrations are employed. Neither BrdU (1–20 μM) nor exposure to black light alone was mutagenic, but the combined treatment did result in the induction of TG^r mutants. The mutant frequency increased with increasing light exposure at constant BrdU and with inreasing BrdU at constant light exposure. These results show that BrdU is incorporated into the hgprt gene, but that this does not result in mutation induction in the absence of light exposure. Such a BrdU-plus-light procedure might be applied to studies of DNA repair at this locus, since mutation induction requires both BrdU incorporation and subsequent exposure to black light.