Changes in DNA 8-hydroxyguanine levels, 8-hydroxyguanine repair activity, and hOGG1 and hMTH1 mRNA expression in human lung alveolar epithelial cells induced by crocidolite asbestos

Carcinogenesis. 2001 Feb;22(2):265-9. doi: 10.1093/carcin/22.2.265.

Abstract

We examined 8-hydroxyguanine (8-OH-Gua) formation and 8-OH-Gua repair enzyme activity in pulmonary type-II-like epithelial cells to determine whether oxidative stress induced by asbestos plays a role in its carcinogenic mechanism. A549 cells were incubated with crocidolite asbestos at concentrations of 0, 10, 50 and 100 microg/ml over 27 h. We then evaluated 8-OH-Gua formation, 8-OH-Gua repair enzyme activity and gene expression of 8-oxoguanine-DNA glycosylase 1 (hOGG1) and human MUtT homologue (hMTH1). This was done using a high-performance liquid chromatography system equipped with an electrochemical detector, endonuclease nicking assay and reverse transcription polymerase chain reaction, respectively. Crocidolite induced the formation of 8-OH-Gua in DNA at concentrations of 50 and 100 microg/ml. 8-OH-Gua levels increased at 9 h and had declined to near baseline at 27 h, whereas 8-OH-Gua repair enzyme activity peaked at 18 h post-crocidolite exposure. hOGG1 and hMTH1 mRNA levels were also increased by crocidolite exposure. These data suggest that crocidolite asbestos is associated with epithelial cell injury in the process of carcinogenesis through oxidative stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Asbestos, Crocidolite / toxicity*
  • DNA Primers / chemistry
  • DNA Repair Enzymes*
  • DNA Repair*
  • DNA-Formamidopyrimidine Glycosylase
  • Endonucleases / metabolism
  • Epithelial Cells / drug effects*
  • Epithelial Cells / metabolism
  • Guanine / analogs & derivatives*
  • Guanine / metabolism*
  • Humans
  • N-Glycosyl Hydrolases / biosynthesis
  • N-Glycosyl Hydrolases / genetics*
  • Oxidative Stress
  • Phosphoric Monoester Hydrolases / biosynthesis
  • Phosphoric Monoester Hydrolases / genetics*
  • Pulmonary Alveoli / drug effects*
  • Pulmonary Alveoli / metabolism
  • RNA, Messenger / biosynthesis*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Superoxide Dismutase / metabolism
  • Tumor Cells, Cultured

Substances

  • Actins
  • DNA Primers
  • RNA, Messenger
  • Asbestos, Crocidolite
  • 8-hydroxyguanine
  • Guanine
  • Superoxide Dismutase
  • Endonucleases
  • Phosphoric Monoester Hydrolases
  • N-Glycosyl Hydrolases
  • DNA-Formamidopyrimidine Glycosylase
  • 8-oxodGTPase
  • DNA Repair Enzymes