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Effect of doxorubicin on selected oxidative stress markers in adenocarcinoma cell line A549

Renata M. Polaniak 1Rafal J. Buldak 2Małgorzata T. Latocha 3Ewa Romuk 1Mateusz Grajek 4Ryszard Szkilnik 5Ewa Birkner 1

1. Department of General Biochemistry, Faculty of Medicine in Zabrze, Silesian Medical University, Jordana 19, Zabrze 41-808, Poland
2. Department of Physiology, Medical University of Silesia, H. Jordana 38, Zabrze 41-808, Poland
3. Department of Cell Biology, Medical University of Silesia (SUM), Jedności 8, Sosnowiec 41-200, Poland
4. Department of Dietetics, Medical Uniwersity of Silesia, H.Jordana 19, Zabrze 41-808, Poland
5. Department of Basic Medical Sciences, Medical Universityof Silesia, Piekarska18, Bytom 41-902, Poland


Doxorubicin - DOX (adriamycin), belongs to the group of anthracycline antibiotics. The molecule of this substance contains aglycone and sugar moiety. Doxorubicin and its anthracycline derivatives are involved in free radical reactions, lipid peroxidation and changes in cellular structures. The mechanism of toxicity of doxorubicin and its anthracycline derivatives relies on a number of free radical reactions that start lipid peroxidation processes in the cell, which in turn can cause damage to cellular structures and functions. Target molecules of the reactive forms of oxygen are, in the first place, DNA, proteins and phospholipids present in the structures of cell membranes. The degree of cell damage by oxidative stress is a result of both its intensity and reduced antioxidant potential. The aim of the study was to demonstrate the impact of doxorubicin, in the used dose and within selected time interval, on cancer cells of the studied line in vitro and to evaluate the dynamics of changes in the activity of enzymes belonging to the main prooxidant/ : superoxide dismutase and its isoenzymes (mitochondrial, MnSOD and cytoplasmic Cu/ZnSOD) and peroxidase glutamate. As a marker of lipid peroxidation, serum malondialdehyde (MDA) was used. Cultures were grown on the A549 adenocarcinoma line. The culture medium was Dulbeco's with L-glutamine, fetal bovine serum enhanced. Breeding was conducted under standard conditions of temperature and carbon dioxide-enriched atmosphere. Cells were treated with three concentrations of doxorubicin: D1=0,125mg; D2 = 0,25mg/ml; D3 = 0,5mg/ml. After 24-hour exposure in cell supernatant, activity of studied enzymes and the concentration of serum malondialdehyde was measured. The research found that doxorubicin in the used doses and within selected time limit interferes with the activity of major oxidative stress systems of cancer cells of the studied line in in-vitro conditions. The effect depends on dose and duration of action of cytostatic drugs. The study also demonstrated a relationship between the chemoresistance of the cancer cells of the line A549 in vitro and the activity of mitochondrial superoxide dismutase isoform.


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Submitted: 2014-03-15 21:24
Revised:   2014-05-02 17:33