Protection of human lung cells against hyperoxia using the DNA base excision repair genes hOgg1 and Fpg.
Hyperoxia causes pulmonary toxicity in part by injuring alveolar epithelial cells. Previous studies have shown that toxic oxygen-derived species damage DNA and this damage is recognized and repaired by either human enzyme 8-oxoguanine DNA glycosylase (hOgg1) or Escherichia coli enzyme formamidopyrimidine DNA glycosylase (Fpg). To determine whether these DNA repair proteins can reduce O(2)-mediated DNA damage in lung cells, A549 lung epithelial cells were transduced with either hOgg1 or Fpg using a retroviral vector containing enhanced green fluorescent protein. Expression of each gene in the transduced cells was confirmed by fluorescent microscopy, Northern blotting, Western blotting, and an enzymatic oligonucleotide cleavage assay. A549 cells expressing either hOgg1 or Fpg were protected from hyperoxia as evidenced by a decrease in DNA damage and a corresponding increase in cell survival. Further, we determined that overexpression of hOgg1 or Fpg partially mitigated the toxic effects of hydrogen peroxide in lung cells. Our data suggest that increased expression of DNA base excision repair genes might represent a new approach for protecting critical lung cells from the toxic effects of hyperoxia.
Wu
Min
M
Division of Pulmonary, Allergy, Critical Care, and Occupational Medicine, Department of Medicine, Indiana University School of Medicine, Indianapolis, 46202, USA.
He
Ying-Hui
YH
Kobune
Masayoshi
M
Xu
Yi
Y
Kelley
Mark R
MR
Martin
William J
WJ
D002470:Cell Survival; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D020552:Comet Assay; D004260:DNA Repair / Q000235:genetics*; D043382:DNA-Formamidopyrimidine Glycosylase; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D018496:Hyperoxia / Q000201:enzymology; D008168:Lung / Q000201:enzymology / Q000503:physiopathology*; D009699:N-Glycosyl Hydrolases / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D010100:Oxygen / Q000502:physiology*; D014161:Transduction, Genetic
D029968:Escherichia coli Proteins; D006861:Hydrogen Peroxide; D009699:N-Glycosyl Hydrolases; D043382:DNA-Formamidopyrimidine Glycosylase; C468610:DNA-formamidopyrimidine glycosylase, E coli; D010100:Oxygen