Molecular mechanisms of the effects of N-ethylmaleimide and 1,4-dithioerythritol on regulation of apoptosis in P19 cells under hypoxia
Relevance. Impairment of apoptosis regulation in P19 cells is correlated with generation of oxidative stress. Under hypoxia, changes in mitochondrial functions occur, which may exacerbate oxidative stress in the tumor cell. The aim of the study was to evaluate the effects of N-ethylmaleimide and 1,4...
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| Format: | Article |
| Language: | English |
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Siberian State Medical University (Tomsk)
2020-07-01
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| Series: | Бюллетень сибирской медицины |
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| Online Access: | https://bulletin.ssmu.ru/jour/article/view/2863 |
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| author | O. L. Nosareva D. S. Orlov E. V. Shakhristova E. A. Stepovaya |
| author_facet | O. L. Nosareva D. S. Orlov E. V. Shakhristova E. A. Stepovaya |
| author_sort | O. L. Nosareva |
| collection | DOAJ |
| description | Relevance. Impairment of apoptosis regulation in P19 cells is correlated with generation of oxidative stress. Under hypoxia, changes in mitochondrial functions occur, which may exacerbate oxidative stress in the tumor cell. The aim of the study was to evaluate the effects of N-ethylmaleimide and 1,4-dithioerythritol on implementation and regulation of apoptosis in P19 cells under hypoxia in vitro.Materials and methods. P19 cells (mouse teratocarcinoma) cultured under hypoxia served as the material for the study. For redox status modulation, 5mM N-ethylmaleimide and 1,4-dithioerythritol in the final concentrations of 5 mM were used. The intracellular concentration of calcium ions, the transmembrane potential and the number of Annexin V, CD95 and CD120 positive cells were determined by flow cytometry. The levels of reduced, oxidized and protein-bound glutathione, protein SH groups, hydroxyl radical and protein carbonyl derivatives were measured by spectrophotometry.Results. The alteration in the redox status of the glutathione system under hypoxia, accompanied by oxidative modification of proteins (glutathionylation and carbonylation), influences the metabolism in the tumor cell on the whole. Under the effects of 1,4-dithioerythritol, an SH group protector, this alteration promotes formation of additional mechanisms to escape apoptosis, whereas under the effects of N-ethylmaleimide, an SH group blocker, it, on the contrary, promotes apoptosis activation.Conclusions. The changes in the redox homeostasis of the tumor cell and modulation of oxidative modification of proteins (glutathionylation and carbonylation) under hypoxia are one of the promising approaches to targeted regulation of cell death. |
| first_indexed | 2024-04-10T01:05:39Z |
| format | Article |
| id | doaj.art-c827426e70c84b519711ea51f83266d8 |
| institution | Directory Open Access Journal |
| issn | 1682-0363 1819-3684 |
| language | English |
| last_indexed | 2024-04-10T01:05:39Z |
| publishDate | 2020-07-01 |
| publisher | Siberian State Medical University (Tomsk) |
| record_format | Article |
| series | Бюллетень сибирской медицины |
| spelling | doaj.art-c827426e70c84b519711ea51f83266d82023-03-13T09:58:26ZengSiberian State Medical University (Tomsk)Бюллетень сибирской медицины1682-03631819-36842020-07-01192727710.20538/1682-0363-2020-2-72-771623Molecular mechanisms of the effects of N-ethylmaleimide and 1,4-dithioerythritol on regulation of apoptosis in P19 cells under hypoxiaO. L. Nosareva0D. S. Orlov1E. V. Shakhristova2E. A. Stepovaya3Сибирский государственный медицинский университет (СибГМУ)Сибирский государственный медицинский университет (СибГМУ)Сибирский государственный медицинский университет (СибГМУ)Сибирский государственный медицинский университет (СибГМУ)Relevance. Impairment of apoptosis regulation in P19 cells is correlated with generation of oxidative stress. Under hypoxia, changes in mitochondrial functions occur, which may exacerbate oxidative stress in the tumor cell. The aim of the study was to evaluate the effects of N-ethylmaleimide and 1,4-dithioerythritol on implementation and regulation of apoptosis in P19 cells under hypoxia in vitro.Materials and methods. P19 cells (mouse teratocarcinoma) cultured under hypoxia served as the material for the study. For redox status modulation, 5mM N-ethylmaleimide and 1,4-dithioerythritol in the final concentrations of 5 mM were used. The intracellular concentration of calcium ions, the transmembrane potential and the number of Annexin V, CD95 and CD120 positive cells were determined by flow cytometry. The levels of reduced, oxidized and protein-bound glutathione, protein SH groups, hydroxyl radical and protein carbonyl derivatives were measured by spectrophotometry.Results. The alteration in the redox status of the glutathione system under hypoxia, accompanied by oxidative modification of proteins (glutathionylation and carbonylation), influences the metabolism in the tumor cell on the whole. Under the effects of 1,4-dithioerythritol, an SH group protector, this alteration promotes formation of additional mechanisms to escape apoptosis, whereas under the effects of N-ethylmaleimide, an SH group blocker, it, on the contrary, promotes apoptosis activation.Conclusions. The changes in the redox homeostasis of the tumor cell and modulation of oxidative modification of proteins (glutathionylation and carbonylation) under hypoxia are one of the promising approaches to targeted regulation of cell death.https://bulletin.ssmu.ru/jour/article/view/2863редокс-статусопухолевый ростокислительный стресссистема глутатионаапоптозгипоксия |
| spellingShingle | O. L. Nosareva D. S. Orlov E. V. Shakhristova E. A. Stepovaya Molecular mechanisms of the effects of N-ethylmaleimide and 1,4-dithioerythritol on regulation of apoptosis in P19 cells under hypoxia Бюллетень сибирской медицины редокс-статус опухолевый рост окислительный стресс система глутатиона апоптоз гипоксия |
| title | Molecular mechanisms of the effects of N-ethylmaleimide and 1,4-dithioerythritol on regulation of apoptosis in P19 cells under hypoxia |
| title_full | Molecular mechanisms of the effects of N-ethylmaleimide and 1,4-dithioerythritol on regulation of apoptosis in P19 cells under hypoxia |
| title_fullStr | Molecular mechanisms of the effects of N-ethylmaleimide and 1,4-dithioerythritol on regulation of apoptosis in P19 cells under hypoxia |
| title_full_unstemmed | Molecular mechanisms of the effects of N-ethylmaleimide and 1,4-dithioerythritol on regulation of apoptosis in P19 cells under hypoxia |
| title_short | Molecular mechanisms of the effects of N-ethylmaleimide and 1,4-dithioerythritol on regulation of apoptosis in P19 cells under hypoxia |
| title_sort | molecular mechanisms of the effects of n ethylmaleimide and 1 4 dithioerythritol on regulation of apoptosis in p19 cells under hypoxia |
| topic | редокс-статус опухолевый рост окислительный стресс система глутатиона апоптоз гипоксия |
| url | https://bulletin.ssmu.ru/jour/article/view/2863 |
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