Protection of pulmonary epithelial cells from oxidative stress by hMYH adenine glycosylase

Ted M. Kremer, Mikael L. Rinne, Yi Xu, Xian Ming Chen, Mark Kelley

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background: Oxygen toxicity is a major cause of lung injury. The base excision repair pathway is one of the most important cellular protection mechanisms that responds to oxidative DNA damage. Lesion-specific DNA repair enzymes include hOgg1, hMYH, hNTH and hMTH. Methods: The above lesion-specific DNA repair enzymes were expressed in human alveolar epithelial cells (A549) using the pSF91.1 retroviral vector. Cells were exposed to a 95% oxygen environment, ionizing radiation (IR), or H2O2. Cell growth analysis was performed under non-toxic conditions. Western blot analysis was performed to verify over-expression and assess endogenous expression under toxic and non-toxic conditions. Statistical analysis was performed using the paired Student's t test with significance being accepted for p < 0.05. Results: Cell killing assays demonstrated cells over-expressing hMYH had improved survival to both increased oxygen and IR. Cell growth analysis of A549 cells under non-toxic conditions revealed cells over-expressing hMYH also grow at a slower rate. Western blot analysis demonstrated over-expression of each individual gene and did not result in altered endogenous expression of the others. However, it was observed that O2 toxicity did lead to a reduced endogenous expression of hNTH in A549 cells. Conclusion: Increased expression of the DNA glycosylase repair enzyme hMYH in A549 cells exposed to O2 and IR leads to improvements in cell survival. DNA repair through the base excision repair pathway may provide an alternative way to offset the damaging effects of O2 and its metabolites.

Original languageEnglish
JournalRespiratory Research
Volume5
DOIs
StatePublished - Sep 27 2004

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Oxidative Stress
Epithelial Cells
DNA Repair Enzymes
Lung
Ionizing Radiation
DNA Repair
Oxygen
Western Blotting
DNA Glycosylases
Alveolar Epithelial Cells
Poisons
Lung Injury
Growth
DNA Damage
adenine glycosylase
Cell Survival
Students
Survival
Genes
A549 Cells

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

Protection of pulmonary epithelial cells from oxidative stress by hMYH adenine glycosylase. / Kremer, Ted M.; Rinne, Mikael L.; Xu, Yi; Chen, Xian Ming; Kelley, Mark.

In: Respiratory Research, Vol. 5, 27.09.2004.

Research output: Contribution to journalArticle

Kremer, Ted M. ; Rinne, Mikael L. ; Xu, Yi ; Chen, Xian Ming ; Kelley, Mark. / Protection of pulmonary epithelial cells from oxidative stress by hMYH adenine glycosylase. In: Respiratory Research. 2004 ; Vol. 5.
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abstract = "Background: Oxygen toxicity is a major cause of lung injury. The base excision repair pathway is one of the most important cellular protection mechanisms that responds to oxidative DNA damage. Lesion-specific DNA repair enzymes include hOgg1, hMYH, hNTH and hMTH. Methods: The above lesion-specific DNA repair enzymes were expressed in human alveolar epithelial cells (A549) using the pSF91.1 retroviral vector. Cells were exposed to a 95{\%} oxygen environment, ionizing radiation (IR), or H2O2. Cell growth analysis was performed under non-toxic conditions. Western blot analysis was performed to verify over-expression and assess endogenous expression under toxic and non-toxic conditions. Statistical analysis was performed using the paired Student's t test with significance being accepted for p < 0.05. Results: Cell killing assays demonstrated cells over-expressing hMYH had improved survival to both increased oxygen and IR. Cell growth analysis of A549 cells under non-toxic conditions revealed cells over-expressing hMYH also grow at a slower rate. Western blot analysis demonstrated over-expression of each individual gene and did not result in altered endogenous expression of the others. However, it was observed that O2 toxicity did lead to a reduced endogenous expression of hNTH in A549 cells. Conclusion: Increased expression of the DNA glycosylase repair enzyme hMYH in A549 cells exposed to O2 and IR leads to improvements in cell survival. DNA repair through the base excision repair pathway may provide an alternative way to offset the damaging effects of O2 and its metabolites.",
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