Evaluation of mutagenic effects of hyperbaric oxygen (HBO) in vitro. II. Induction of oxidative DNA damage and mutations in the mouse lymphoma assay.
We recently showed that treatment of V79 cells with hyperbaric oxygen (HBO) efficiently induced DNA effects in the comet assay and chromosomal damage in the micronucleus test (MNT), but did not lead to gene mutations at the hprt locus. Using the comet assay in conjunction with bacterial formamidopyrimidine DNA glycosylase (FPG protein), we now provide indirect evidence that the same treatment leads to the induction of 8-oxoguanine, a premutagenic oxidative DNA base modification in V79 and mouse lymphoma (L5178Y) cells. We also demonstrate that HBO efficiently induces mutations in the mouse lymphoma assay (MLA). Exposure of L5178Y cells to HBO (98% O(2); 3bar) for 2h caused a clear mutagenic effect in the MLA, which was further enhanced after a 3h exposure. As this mutagenic effect was solely due to the strong increase of small colony (SC) mutants, we suggest that HBO causes mutations by induction of chromosomal alterations. Molecular characterization of induced SC mutants by loss of heterozygosity (LOH) analysis showed an extensive loss of functional tk sequences similar to the pattern found in spontaneous SC mutants. This finding confirmed that the majority of HBO-induced mutants is actually produced by a clastogenic mechanism. The induction of point mutations as a consequence of induced oxidative DNA base damage seems to be of minor importance.
Rothfuss
A
A
Universitätsklinikum Ulm, Abteilung Humangenetik, D-89070 Ulm, Germany.
Merk
O
O
Radermacher
P
P
Speit
G
G
D000818:Animals; D002460:Cell Line; D020552:Comet Assay; D006224:Cricetinae; D003412:Cricetulus / Q000235:genetics; D004249:DNA Damage / Q000187:drug effects*; D004273:DNA, Neoplasm / Q000032:analysis; D043382:DNA-Formamidopyrimidine Glycosylase; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D006147:Guanine / Q000031:analogs & derivatives* / Q000096:biosynthesis; D006931:Hyperbaric Oxygenation / Q000009:adverse effects*; D007940:Leukemia L5178 / Q000201:enzymology / Q000235:genetics*; D019656:Loss of Heterozygosity; D009152:Mutagenicity Tests; D009153:Mutagens / Q000633:toxicity*; D009699:N-Glycosyl Hydrolases / Q000235:genetics; D010100:Oxygen / Q000633:toxicity*; D017354:Point Mutation / Q000187:drug effects*; D016133:Polymerase Chain Reaction; D013937:Thymidine Kinase / Q000235:genetics*
D004273:DNA, Neoplasm; D009153:Mutagens; C024829:8-hydroxyguanine; D006147:Guanine; D013937:Thymidine Kinase; D009699:N-Glycosyl Hydrolases; D043382:DNA-Formamidopyrimidine Glycosylase; D010100:Oxygen