Induction of oxidative stress by hydrogen peroxide in vitro in breast epithelial cells for studying MCF-7 tumor cell line apoptosis
Author Affiliations1Siberian State Medical University, 634050, Tomsk, Moscowski Trakt, 2
2Siberian State Medical University of Minzdrav of Russia, 634050, Tomsk, Moskovsky Tract, 2; National Research Tomsk State University, 634050, Tomsk, Lenin av., 36
3Siberian State Medical University, 634050, Tomsk, Moscowski Trakt, 2
4Siberian State Medical University, 634050, Tomsk, Moscowski Trakt, 2
Abstract
The research objective. Selection of the hydrogen peroxide concentration that is able to induce oxidative stress and trigger breast epithelial cell apoptosis without eliminating cells via cell death. Material and methods. The cell culture of human breast epithelium (HBL-100) and MCF-7 tumor cell line (epithelial-like eadenocarcinoma of human breast) have been chosen as the objects of the research. Modeling of oxidative stress in HBL-100 cell line has been achieved by adding hydrogen peroxide in the final concentrations of 0.2; 0.3; 0.5 and 1.0 mMole. The level of reactive oxygen species, the concentration of total, reduced and oxidized glutathione as well as their ratio in HBL-100 epithelial cell line have been evaluated in order to carry out comparative analysis of the degree of oxidative stress with MCF-7 tumor cells line. Results and discussion. The optimal concentration of hydrogen peroxide has been established – 0.3 mMole, at which the increase in the concentration of reactive oxygen species in the cell is registered that may be compared to the one in tumor cells and also may be compared to the value of the reduced glutathione-oxidized glutathione ratio; more over, formation of the maximum amount of annexin V-positive cells and the minimum amount of propidium iodide- positive cells takes place.
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References
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About Authors (Correspondence):
Shakhristova E.V. – candidate of medical sciences, head of the laboratory of molecular medicine
Ryazantseva N.V. – doctor of medical sciences, professor, head of the chair for molecular medicine and clinical laboratory diagnostics, e-mail: office@ssmu.tomsk.ru
Stepovaya E.A. – doctor of medical sciences, professor of the chair for biochemistry and molecular biology, e-mail: muir@mail.ru
Nosareva О.L. – candidate of medical sciences, assistant professor of the chair for biochemistry and molecular biology, e-mail: olnosareva@yandex.ru
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