№ 2 / 2016 / 19-23

The role of glutathione and protein oxidative modification in apoptosis of breast epithelial cells under induced oxidative stress

Author: Shakhristova E.V.¹, Stepovaya E.A.², Ryazantseva N.V.³, Nosareva О.L.⁴, Chil’chigashev R.I.⁵, Novitsky V.V.⁶

Abstract

The research objective. The aim of the study was to establish the role of glutathione and proteins oxidative modifications in the breast cells apoptosis realization during cultivation with hydrogen peroxide. Material and methods. The cell culture of human breast epithelium (HBL-100) has been chosen as the objects of the research. Oxidative stress in cells was induced by hydrogen peroxide 0.3 mMole. A protector of protein SH groups, 1.4-dithioerythritol, was used to modulate redox status of HBL-100 cells under the conditions of free radical oxidation activation. The concentration of intracellular reactive oxygen species, protein carbonyl derivatives, reduced and oxidized glutathione as well as the intensity of apoptotic cell death were evaluated under the conditions of oxidative stress and HBL-100 cell culturing in the presence of 1.4-dithioerythritol. Results and discussion. It was found that 1,4-dithioerythritol has protective effect on cell proteins, preventing their irreversible carbonylation by restoring glutathione redox potential. Moreover, it reduces apoptosis of breast epithelial cells under oxidative stress.

Key words

apoptosis, сarbonyl derivatives of proteins, breast epithelial cell, oxidative stress, cell redox status

References

Arutjunjan A.V., Dubinina E.E., Zybina N.N. Methods of assessing free radical oxidation and antioxidant protection of the organism. St. Petersburg, 2000. 104 р. [In Russian]
Dubininа E.E. Products of metabolism of oxygen in the functional activity of cells (life and death, creation and destruction). Physiological and clinical-biochemical aspects. Saint-Petersburg, 2006. 400 р. [In Russian]
Kondakova I.V., Kakurina G.V., Smirnova L.P., Borunov E.V. Regulation of tumor cell proliferation and apoptosis by free radicals // Siberian journal of oncology. 2005. (1). 58–61. [In Russian]
Kulinskij V.I., Kolesnichenko L.S. Glutathione system. I. Synthesis, transport, glutathione transferase, glutathione peroxidase. Biomeditsinskaya Khimiy. 2009. 55. (3). 255–277. [In Russian]
Menshchikova E.B., Zenkov N.K., Lankin V.Z. et al. Oxidative stress: Pathological conditions and diseases. Novosibirsk, 2008. 284р. [In Russian]
Oktyabrsky O.N., Smirnova C.V. Redox regulation of cellular functions // Biochemistry. 2007. 72. (2). 158–174. [In Russian]
Stepovaya E.A., Petina G.V., Zhavoronok T.V. et al. Role of thiol-disulfide system in mechanisms of functional changes in neutrophils under conditions of oxidative stress // Bull. Exp. Biol. Med. 2010. 150. (2). 198–202. [In Russian]
Shakhristova E.V., Ryazantseva N.V., Stepovaya E.A., Nosareva O.L. Induction of oxidative stress by hydrogen peroxide in vitro in breast epithelial cells for studying MCF-7 tumor cell line apoptosis // Siberian scientific medical journal. 2015. 35. (2). 5–10. [In Russian]
Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding // Anal. Biochem. 1976. 7. (1, 2). 248–254.
Brigelius-Flohé R., Flohé L. Basic principles and emerging concepts in the redox control of transcription factors // Antioxid. Redox Signal. 2011. 15. (8). 2335–2381.
Brunelli L., Crow J.P., Beckman J.S. The comparative toxicity of nitric oxide and peroxynitrite to Escherichia coli // Arch. Biochem. Biophys. 1995. 316. (1). 327–333.
Halliwell B. Biochemistry of oxidative stress // Biochem. Soc. Trans. 2007. (5). 1147–1150.
Halliwell B., Whiteman M. Measuring reactive species and oxidative damage in vivo and in cell culture: how should you do it and what do the results mean? // Br. J. Pharmacol. 2004. (142). 231–255.
Rahman I., Kode A., Biswas S.K. Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method // Nat. Protoc. 2006. 1. (6). 3159–3165.
Wong C.M., Bansal G., Marcocci L., Suzuki Y.J. Proposed role of primary protein carbonylation in cell signaling // Redox Rep. 2012. 17 (2). 90–94.

About Authors (Correspondence):

Shakhristova E.V. – candidate of medical sciences, head of the laboratory of molecular medicine, assistant professor of the chair for biochemistry and molecular biology with the course of clinical laboratory diagnostics, e-mail: shaxristova@yandex.ru

Stepovaya E.A. – doctor of medical sciences, professor of the chair for biochemistry and molecular biology with the course of clinical laboratory diagnostics, e-mail: muir@mail.ru

Ryazantseva N.V. – doctor of medical sciences, professor of the chair for biophysics, professor of the chair for biological chemistry with courses of medical, pharmaceutical and toxicological chemistry

Nosareva О.L. – candidate of medical sciences, assistant professor of the chair for biochemistry and molecular biology with the course of clinical laboratory diagnostics, e-mail: olnosareva@yandex.ru

Chil’chigashev R.I. – student of the 5th course of medical biological faculty

Novitsky V.V. – doctor of medical sciences, professor, academician of RAS, head of the chair for pathophysiology

Full Text

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Received: 19/05/2016