STUDIES ON INTRACHROMOSOMAL RECOMBINATION IN SP5/V79 CHINESE-HAMSTER CELLS UPON EXPOSURE TO DIFFERENT AGENTS RELATED TO CARCINOGENESIS

In the present study 34 agents, related to carcinogenesis in different ways, were investigated with respect to their recombinogenic activity in mammalian cells. The induction of intrachromosomal recombination was studied using the spontaneous mutant clone SP5 derived from V79 Chinese hamster cells, which exhibits a duplication of exon 2 and its flanking regions in the hprt gene, which was found to be inserted between the two EcoR1 sites of intron 1. Earlier studies on the removal of this insertion fragment in the SP5 clone indicated that such loss involved intrachromosomal recombination and was detectable by using a reversion mutation assay. The categories of agents investigated here included monofunctional alkylating agents, polyaromatic hydrocarbons giving rise to bulky adducts, chlorinated compounds giving small cyclic adducts, intercalating agents, DNA cross-linkers, UV and ionizing radiation, inhibitors of DNA synthesis and topoisomerases, DNA bases and base analogues, radical formers and tumour promoters. Statistically significant enhancements in the frequency of reversion in SP5 cells were observed after treatment with aflatoxin B-1, 9-aminoacridine, benzo[a]pyrene-7,8-dihydrodiol, benzo[a]pyrene-7,8-diol-9,10-epoxide, camptothecin, dimethylbenzanthracene, dimethylnitrosamine, ethidium bromide, ethylmethanesulfonate, N-ethyl-N'-nitrosourea, fluorodeoxyuridine, ICR 191, N-methyl-N'-nitrosoguanidine, mitomycin C and UV irradiation. Only slight inducing effects were indicated in the case of methylmethanesulfonate, N-methyl-N' -nitrosourea and gamma irradiation, although not statistically significant. Negative results were found after treatment with 3-aminobenzamide, 5-azacytidine, bleomycin, 5-bromodeoxyuridine, 1,2-dichloroethane, ethylene oxide, etoposide, formaldehyde, hydrogen peroxide, methotrexate, propylene oxide, quercitin, sodium azide, 12-O-tetradecanoylphorbol-13-acetate, thymidine and a complex mixture consisting of a cigarette smoke condensate. Our results on chemically or physically induced recombinogenic effects in the endogenous SP5 hprt gene are in agreement with data obtained in non-endogenous gene systems based on transgenic cell lines containing integrates of tandem mutated tk or hygromycin resistance genes, but not completely consistent with findings on integrates based on the neo genes. This suggests that many factors influence the recombination process, including a difference in the mechanisms underlying inter- and intrachromosomal recombination. Consequently, the endogenous SP5/V79 system is suggested to be more representative than integrated systems for investigating induction of recombination, as well as for mechanistic studies of recombination at the molecular level, e.g. intrachromosomal recombination.