Tag Archives: ROS

Adaptor protein Ruk/CIN85 affects redox balance in breast cancer cells

I. R. Horak*, N. V. Latyshko, O. O. Hudkova, T. O. Kishko,
O. V. Khudiakova, D. S. Gerashchenko, T. D. Skaterna,
I. P. Krysiuk, S. G. Shandrenko, L. B. Drobot

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: iryna.horak@gmail.com

Received: 25 February 2020; Accepted: 15 May 2020

Excessive reactive oxygen species (ROS) production may lead to damage of cellular proteins, lipids and DNA, and cause cell death. Our previous findings demonstrated that increased level of adaptor protein Ruk/CIN85 contributes to breast cancer cells malignancy. The aim of this study was to investigate the role of Ruk/CIN85 in the maintaining of the redox balance in cancer cells. Mouse breast adenocarcinoma 4T1 cells with different levels of Ruk/CIN85 expression were used as a model in this study. Activities of catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), aldehyde dehydrogenase (ALDH) and formaldehyde dehydrogenase (FALDH), as well as H2O2 and aldehydes content were measured using fluorometric assays. Gene expression correlations between Ruk/CIN85 and antioxidant enzymes in breast cancer samples were analyzed using ist.medisapiens transcriptomic database. It was demonstrated that Ruk/CIN85-overexpressing 4T1 cells were characterized by increased production of H2O2 and reduced activities of CAT, GPx and SOD. Overexpression of Ruk/CIN85 resulted in decreased content of aldehydes together with increased activity of ALDH, while in Ruk/CIN85-knocked down 4T1 cells, activities of ALDH and FALDH were decreased. The data of transcriptomic analysis revealed the correlations between SH3KBP1 expression and CAT, GPX4, ALDH1A1, ALDH1L1, ALDH2, GSR, SOD1 in human breast carcinomas samples. The obtained results indicate that adaptor protein Ruk/CIN85 affects redox balance in mouse breast adenocarcinoma 4T1 cells.

Modulation of cisplatin-induced reactive oxygen species production by fullerene C(60) in normal and transformed lymphoid cells

D. V. Franskevych, I. I. Grynyuk, S. V. Prylutska, O. P. Matyshevska

Taras Shevchenko National University of Kyiv, Ukraine;
е-mail: dashaqq@gmail.com

The early response of normal (Wistar rat thymocytes) and transformed (mice lymphoid leukemia L1210) cells to treatment with anticancer drug cisplatin or to combined treatment with cisplatin and carbon nanostructure fullerene C60 was studied. We demonstrated with fluorescent probes DCFH-DA and TMRE that cisplatin at concentration 1 μg/ml induced reactive oxygen species (ROS) production and decreased the value of mitochondrial membrane potential in both cell types. The combined treatment with cisplatin (1 μg/ml) and fullerene C60 (7.2 μg/ml) was shown to be followed by oppositely directed modulation of ROS production in thymocytes and L1210 cells. Cisplatin-induced ROS production was intensified in L1210 cells, while in thymocytes it was decreased. It is supposed that the different effects of combined treatment are associated with peculiarities of fullerene C60 accumulation and localization in normal and cancer cells.

Computer prediction of biological activity of dimethyl-N-(benzoyl)amidophosphate and dimethyl-N-(phenylsulfonyl)amidophosphate, evaluation of their cytotoxic activity against leukemia cells in vitro

I. I. Grynyuk, S. V. Prylutska, N. S. Kariaka, T. Yu. Sliva,
O. V. Moroz, D. V. Franskevych, V. M. Amirkhanov,
O. P. Matyshevska, M. S. Slobodyanik

Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: igrynyuk@yahoo.com

Structural analogues of β-diketones – dimethyl-N-(benzoyl)amidophosphate (HCP) and dimethyl-N-(phenylsulfonyl)amidophosphate (HSP) were synthesized and identified by the methods of IR, 1H and 31P NMR spectroscopy. Screening of biological activity and calculation of physicochemical parameters of HCP and HSP compounds were done with the use of PASS and ACD/Labs computer programs. A wide range of biological activity of synthesized compounds, antitumor activity in particular, has been found. Calculations of the bioavailability criteria indicate that the investigated compounds have no deviations from Lipinski’s rules. HCP compound is characterized by a high lipophilicity at physiological pH as compared to HSP. It was found that cytotoxic effect of the studied compounds on the leukemiс L1210 cells was of time- and dose-dependent character. HCP is characterized by more pronounced and early cytotoxic effects as compared to HSP. It was shown that 2.5 mM HCP increased ROS production 3 times in the early period of incubation, and decreased cell viability by 40% after 48 h, and by 66% – after 72 h. Based on the computer calculation and undertaken research, HCP was selected for target chemical modifications and enhancement of its antitumor effect.

Reactive oxygen species in signal transduction

L. B. Drobot1, A. A. Samoylenko1, A. V. Vorotnikov2, P. A. Tyurin-Kuzmin2,
A. V. Bazalii1, T. Kietzmann3, V. A. Tkachuk2, S. V. Komisarenko1

1Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: drobot@biochem.kiev.ua
2Lomonosov Moscow State University, Faculty of Basic Medicine, Russia;
3Department of Biochemistry and Biocenter Oulu, University of Oulu, Oulu, Finland;

Reactive oxygen species (ROS) are products of incomplete reduction of oxygen both nonradicals and radicals that function as mediators of redox signaling and oxidative stress depending on their levels in different­ subcellular compartments. Up to date, a huge body of data are accumulated, which supports a role of ROS as “second messengers” in intracellular signaling cascades that control cell growth, proliferation, apoptosis as well as migration and invasion. The current review summarizes data regarding ROS-dependent regulation of signaling­ networks components including MAPK, PI3K/Akt, PKC, NF-κB, Nrf2, FoxO and HIF-1α, and role of ROS in tumorigenesis.

Vitamin D(3) availability and functional activity of peripheral blood phagocytes in experimental type 1 diabetes

D. О. Labudzynskyi, І. О. Shymanskyy, V. М. Riasnyi, М. М. Veliky

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: konsument3@gmail.com

The study was devoted to identifying the relation between vitamin D3 availability (assessed by the level of circulatory 25OHD3), content of vitamin D3 25-hydroxylase isozymes CYP27A1 and CYP2R1 in hepatic tissue and functional activity of peripheral blood phagocytes in mice with experimental type 1 diabetes. It has been shown that diabetes is accompanied by the development of vitamin D3-deficiency which is characterized by decreased 25OHD3 content in blood serum and determined by changes in tissue expression of the major isoforms of vitamin D3 25-hydroxylase. The level of hepatic CYP27A1 was revealed to be markedly reduced with a concurrent significant augmentation of CYP2R1. Cholecalciferol administration resulted in normalization of tissue levels of both isoforms of vitamin D3 25-hydroxylase and blood serum 25OHD3 content. Diabetes-associated vitamin D3 deficiency correlated with a decrease in phagocytic activity of granulocytes and monocytes, and their ability to produce antibacterial biooxidants such as reactive oxygen and nitrogen forms. Vitamin D3 efficacy to attenuate these abnormalities of immune function was established, indicating an important immunoregulatory role of cholecalciferol in the phagocytic mechanism of antigens elimination implemented by granulocytes and monocytes.