Primary research

Dissociation of DNA damage and mitochondrial injury caused by hydrogen peroxide in SV-40 transformed lung epithelial cells

Yoshihiro Fujii¹ , Katsuyuki Tomita¹ , Hiroyuki Sano¹ , Akira Yamasaki¹ , Yutaka Hitsuda¹ , Ian M Adcock² and Eiji Shimizu¹

¹  Third Department of Internal Medicine, Faculty of Medicine, Tottori University, 36-1 Nishi-machi, Yonago-shi, Tottori-ken 683-8504, JAPAN

²  Department of Thoracic Medicine, National Heart & Lung Institute at Imperial College School of Science, Technology and Medicine, London, UK

author email corresponding author email

Cancer Cell International 2002, 2:16doi:10.1186/1475-2867-2-16

Published:	20 November 2002

Abstract

Background

Since lung epithelial cells are constantly being exposed to reactive oxygen intermediates (ROIs), the alveolar surface is a major site of oxidative stress, and each cell type may respond differently to oxidative stress. We compared the extent of oxidative DNA damage with that of mitochondrial injury in lung epithelial cells at the single cell level.

Result

DNA damage and mitochondrial injury were measured after oxidative stress in the SV-40 transformed lung epithelial cell line challenged with hydrogen peroxide (H₂O₂). Single cell analysis of DNA damage was determined by assessing the number of 8-oxo-2-deoxyguanosine (8-oxo-dG) positive cells, a marker of DNA modification, and the length of a comet tail. Mitochondrial membrane potential, ΔΨm, was determined using JC-1. A 1 h pulse of H₂O₂ induced small amounts of apoptosis (3%). 8-oxo-dG-positive cells and the length of the comet tail increased within 1 h of exposure to H₂O₂. The number of cells with reduced ΔΨm increased after the addition of H₂O₂ in a concentration-dependent manner. In spite of a continual loss of ΔΨm, DNA fragmentation was reduced 2 h after exposure to H₂O₂.

Conclusion

The data suggest that SV-40 transformed lung epithelial cells are resistant to oxidative stress, showing that DNA damage can be dissociated from mitochondrial injury.