Oxidative damage of U937 human leukemic cells caused by hydroxyl radical results in singlet oxygen formation

PloS One
Marek RácPavel Pospísil

Abstract

The exposure of human cells to oxidative stress leads to the oxidation of biomolecules such as lipids, proteins and nuclei acids. In this study, the oxidation of lipids, proteins and DNA was studied after the addition of hydrogen peroxide and Fenton reagent to cell suspension containing human leukemic monocyte lymphoma cell line U937. EPR spin-trapping data showed that the addition of hydrogen peroxide to the cell suspension formed hydroxyl radical via Fenton reaction mediated by endogenous metals. The malondialdehyde HPLC analysis showed no lipid peroxidation after the addition of hydrogen peroxide, whereas the Fenton reagent caused significant lipid peroxidation. The formation of protein carbonyls monitored by dot blot immunoassay and the DNA fragmentation measured by comet assay occurred after the addition of both hydrogen peroxide and Fenton reagent. Oxidative damage of biomolecules leads to the formation of singlet oxygen as conformed by EPR spin-trapping spectroscopy and the green fluorescence of singlet oxygen sensor green detected by confocal laser scanning microscopy. It is proposed here that singlet oxygen is formed by the decomposition of high-energy intermediates such as dioxetane or tetroxide formed by oxidative da...Continue Reading

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Citations

Feb 13, 2016·Journal of Pharmaceutical Sciences·Olivier MozziconacciChristian Schöneich
Apr 12, 2015·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Mohammad IbrahimJoao Batista Teixeira Rocha
Oct 17, 2019·Journal of Clinical Laboratory Analysis·Bochra GargouriSaloua Lassoued
Dec 6, 2017·Journal of Biophotonics·Hua Shi, Qingming Luo
Nov 13, 2020·Frontiers in Physiology·Ankush PrasadPavel Pospíšil

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Methods Mentioned

BETA
MDA
X-ray
Electrophoresis
dot blot
fluorescence microscopy
carbonylation

Software Mentioned

Winsim
TriTek CometScore Freeware
Statistika

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