title: Dissociation of DNA damage and mitochondrial injury caused by hydrogen peroxide in SV-40 transformed lung epithelial cells.
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, DeltaPsim, 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 DeltaPsim increased after the addition of H2O2 in a concentration-dependent manner. In spite of a continual loss of DeltaPsim, 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.
authors.affiliation: Third Department of Internal Medicine, Faculty of Medicine, Tottori University, 36-1 Nishi-machi, Yonago-shi, Tottori-ken 683-8504, JAPAN. tom0223@grape.med.tottori-u.ac.jp
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Fujii
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Tomita
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Sano
authors.forename: Akira
authors.initials: A
authors.lastname: Yamasaki
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Hitsuda
authors.forename: Ian M
authors.initials: IM
authors.lastname: Adcock
authors.forename: Eiji
authors.initials: E
authors.lastname: Shimizu