Mutation frequency analysis of mononucleotide and dinucleotide repeats after oxidative stress.
Many tumors exhibit genetic instability at the DNA sequence level in the form of frameshift mutations in simple repeats (microsatellite instability). A high level of microsatellite instability, such as that seen in hereditary nonpolyposis colorectal cancer (HNPCC), arises from defects in the mismatch repair pathway. A low level of microsatellite instability is found in some non-HNPCC-associated cancers, such as those of the breast and lung, and is not attributable to mismatch repair defects. We hypothesized that oxidative DNA damage may be at least partly responsible for the generation of microsatellite mutations in these tumors. We investigated whether oxidative DNA damage can induce microsatellite mutations in mismatch repair-proficient cultured cells. Telomerase-immortalized normal human fibroblasts were stably transfected with a plasmid containing a tk-neo fusion gene, such that the neo coding region was placed out of frame by the presence of an upstream microsatellite sequence. Cells were treated with H(2)O(2) and mutation frequencies were determined for G(17), A(17), and (CA)(17) repeats. Mutation frequencies of mononucleotide repeats in cells with the neo gene in the (+1) reading frame were reduced after treatment. No effect was observed in cells with the mononucleotide repeats in the (-1) reading frame. A small increase in mutation frequency was observed in cells with the (CA)(17) repeat. Our data suggest that diploid human cells may have protective mechanisms that prevent the induction of microsatellite mutations by a short exposure to high levels of oxidative stress.
Yamada
Nazumi A
NA
Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
Parker
Jennifer M
JM
Farber
Rosann A
RA
D002461:Cell Line, Transformed; D004249:DNA Damage; D004260:DNA Repair; D005347:Fibroblasts; D016368:Frameshift Mutation; D006801:Humans; D019868:Kanamycin Kinase; D018895:Microsatellite Repeats; D018384:Oxidative Stress; D010957:Plasmids
D019868:Kanamycin Kinase