350 rub
Journal №10 for 2011 г.
Article in number:
Expression of Genes Controlled Biosynthesis of Glutathione under Development of Drug Resistance of Cancer Cells to Doxorubicin
Keywords: drug resistance of cancer cells expression of genes synthesis of glutathione doxorubicin
Authors:
E.V. Kalinina, T.T. Berozov, N.N. Chernov, A.A. Shtil, V.A. Glasunova, M.D. Novichkova, N.K. Nurmuradov
Abstract:
Investigation of a role of redox-regulation, cellular redox state and activity of free radical processes has actual significance for decision of problems connected with the search of courses to overcome restrictions chemotherapy efficiency of tumors by antitumor drugs with pro-oxidant action. The aim of the present research was the study of expression of genes controlled synthesis of glutathione (GSH). The cell lines were used in the study: human erythroleukemia K562, human breast carcinoma MCF-7 and human ovarian carcinoma SKOV-3 - DOX-sensitive cells (K562/S, MCF-7/S, SKOV-3 with IC50 - 0,35, 1,1, 0,2 µМ respectively) and DOX-resistant cells (K562/DOX, MCF-7/DOX, SKVLB with IC50 - 5,2, 25, 1,6 µМ, respectively). mRNA was determined using qRT-PCR. Electrophoresis of PCR products was conducted in 1,5-2% agarose gel followed by densitometry. Gel analysis was performed using the BioCaptMW software (Vilber Lourmat). Activity of γ-glutamylcysteine synthetase (γ-GCS), γ-glutamyltransferase (γ-GT), glutathione reductase (GR), maintenance of GSH and its oxidized form (GSSG), were estimated by spectrophotometry assays. Growth of expression of genes encoding light (γ-GCSL) and heavy (γ-GCSH) subunits of γ-glutamylcysteine synthetase - enzyme limited the rate of GSH biosynthesis and catalyzed formation of γ-glutamyltcysteine, which was co-ordinated with elevation of GS gene controlled GSH formation from γ-glutamyltcysteine and glycine in resistant cells K562/DOX, MCF-7/DOX, SKVLB was found. This effect was suggested by the rise of expression of γ-GT gene controlled transport of γ-glutamyl groups as well as intracellular concentration of cysteine which limits GSH synthesis. Co-ordinative enhanced expression of γ-GCSL, γ-GCSH, GS, γ-GT genes can be estimated as a result of development of optimal conditions for increase of GSH synthesis de novo and together with elevation of expression of gene encoding glutathione reductase causes a growth of intracellular GSH level. The data are indicated enhancement of the role of GSH-dependent processes as a result of development of cancer cells resistance to doxorubicin.
Pages: 35-41
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