Expression of yeast apurinic/apyrimidinic endonuclease (APN1) protects lung epithelial cells from bleomycin toxicity

Y. H. He, M. Wu, M. Kobune, Y. Xu, Mark Kelley, W. J. Martin

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Bleomycin is a well-established anti-tumor drug. Its major un-toward effect, pulmonary toxicity, has limited its usage. In this study, we used a DNA repair protein, yeast apurinic/apyrimidinic endonuclease (APN1) to reduce the toxicity of bleomycin on lung cells. A549 cells, an alveolar epithelial cell line, were transduced by MIEG3 retroviral vector encoding both enhanced green fluorescent protein (EGFP) and APN1. Transduced cells were sorted by fluorescent-activated cell sorter (FACS) analysis and were cloned. The APN1 expression of transduced A549 cell population and four selected clones expressing different levels of EGFP was confirmed by Northern, Western, and apurinic/apyrimidinic (AP) endonuclease activity analyses. The expression of APN1 was positively correlated with the expression of EGFP. The protective effect of APN1 against bleomycin was determined by single cell gel electrophoresis/Comet assay and by clonogenic survival assay following bleomycin treatment. The A549 population expressing APN1 showed a significant reduction of DNA damage in the presence of 20, 50, and 100 μg/ml bleomycin; similarly, the APNl-expressing A549 population also demonstrated increased survival in the presence of bleomycin compared with the vector-transduced A549 population. In selected clones, three of four APN1-expressing clones resulted in significantly improved cell survival. The current study suggests that the yeast DNA repair protein, APN1, can reduce bleomycin toxicity to target lung cells.

Original languageEnglish
Pages (from-to)692-698
Number of pages7
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume25
Issue number6
StatePublished - 2001

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DNA-(Apurinic or Apyrimidinic Site) Lyase
Endonucleases
Bleomycin
Yeast
Toxicity
Yeasts
Epithelial Cells
Lung
Comet Assay
Clone Cells
DNA Repair
Population
Assays
DNA
Repair
Cells
Alveolar Epithelial Cells
Fungal Proteins
Electrophoresis
DNA Damage

ASJC Scopus subject areas

  • Cell Biology
  • Pulmonary and Respiratory Medicine
  • Molecular Biology

Cite this

Expression of yeast apurinic/apyrimidinic endonuclease (APN1) protects lung epithelial cells from bleomycin toxicity. / He, Y. H.; Wu, M.; Kobune, M.; Xu, Y.; Kelley, Mark; Martin, W. J.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 25, No. 6, 2001, p. 692-698.

Research output: Contribution to journalArticle

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