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1 January 2005 Multiplex Polymerase Chain Reaction Analysis of UV-A– and UV-B–induced Delayed and Early Mutations in V79 Chinese Hamster Cells
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We previously reported that approximately 10% of V79 Chinese hamster fibroblast populations clonally derived from single cells immediately after irradiation with either ultraviolet B (UV-B, 290–320 nm, mainly 311 nm) or ultraviolet A (UV-A, 320–400 nm, mainly 350–390 nm) radiation exhibit genomic instability. The instability is revealed by relatively high mutation frequencies in the hypoxanthine phosphoribosyl transferase (hprt) gene up to 23 cell generations after irradiation. These delayed mutant clones exhibited higher levels of oxidative stress than normal cells. Therefore, persistently increased oxidative stress has been proposed as a mechanism for UV-induced genomic instability. This study investigates whether this mechanism is reflected in the deletion spectrum of delayed mutant clones. Eighty-eight percent of the delayed mutant clones derived from UV-A–irradiated populations were found to have total deletion of the hprt gene. Correspondingly, 81% of UV-A–induced early mutations (i.e. detected shortly after irradiation) also had total deletions. Among delayed UV-B–induced mutant clones, 23% had total deletions and 8% had deletion of one exon, whereas all early UV-B events were either point mutations or small deletions or insertions. In conclusion, the multiplex polymerase chain reaction deletion screen showed that there were explicit differences in the occurrence of large gene alterations between early and delayed mutations induced by UV-B radiation. For UV-A radiation the deletion spectra were similar for delayed and early mutations. UV-A radiation is, in contrast to UV-B radiation, only weakly absorbed by DNA and probably induces mutation almost solely via production of reactive oxygen species. Therefore, the present results support the hypothesis that persistent increase in oxidative stress is involved in the mechanism of UV-induced genomic instability.

Jostein Dahle, Paul Noordhuis, Trond Stokke, Debbie Hege Svendsrud, and Egil Kvam "Multiplex Polymerase Chain Reaction Analysis of UV-A– and UV-B–induced Delayed and Early Mutations in V79 Chinese Hamster Cells," Photochemistry and Photobiology 81(1), 114-119, (1 January 2005).
Received: 19 May 2004; Accepted: 1 September 2004; Published: 1 January 2005

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