Kotsarenko Kateryna
@jcu.cz
University of South Bohemia in v Ceske Budejovice
Scopus Publications
Scopus Publications
Kateryna Kotsarenko, Pavlina Vechtova, Jaroslava Lieskovska, Zoltán Füssy, Diogo C. Cabral-de-Mello, Ryan O. M. Rego, Pilar Alberdi, Marisol Collins, Lesley Bell-Sakyi, Jan Sterba,et al.
Springer Science and Business Media LLC
Tick cell lines are an easy-to-handle system for the study of viral and bacterial infections and other aspects of tick cellular processes. Tick cell cultures are often continuously cultivated, as freezing can affect their viability. However, the long-term cultivation of tick cells can influence their genome stability. In the present study, we investigated karyotype and genome size of tick cell lines. Though 16S rDNA sequencing showed the similarity between Ixodes spp. cell lines at different passages, their karyotypes differed from 2n = 28 chromosomes for parental Ixodes spp. ticks, and both increase and decrease in chromosome numbers were observed. For example, the highly passaged Ixodes scapularis cell line ISE18 and Ixodes ricinus cell lines IRE/CTVM19 and IRE/CTVM20 had modal chromosome numbers 48, 23 and 48, respectively. Also, the Ornithodoros moubata cell line OME/CTVM22 had the modal chromosome number 33 instead of 2n = 20 chromosomes for Ornithodoros spp. ticks. All studied tick cell lines had a larger genome size in comparison to the genomes of the parental ticks. Thus, highly passaged tick cell lines can be used for research purposes, but possible differences in encoded genetic information and downstream cellular processes, between different cell populations, should be taken into account.
Kateryna Kotsarenko, Pavlina Vechtova, Zuzana Hammerova, Natalia Langova, Lenka Malinovska, Michaela Wimmerova, Jan Sterba, and Libor Grubhoffer
Elsevier BV
DNA methylation at the fifth position of cytosine (5mC) and at the sixth position of adenine (6 mA) plays an important role in the regulation of the gene expression and, in eukaryotes, is essential for normal development. For Ixodes ricinus, the most common European arthropod vector of human and animal pathogens, the DNA methylation profile and the role of DNA methylation in tick development are still under discussion. Our goal was to analyze the status of I. ricinus DNA methylation at different life stages and identify enzymes that produce this type of DNA modification. We found that 5mC and 6mA are present in I. ricinus genomic DNA at all life stages. In the transcriptome of I. ricinus, we identified the sequences of the putative IrDNMT1, IrDNMT3, and IrDAMT enzymes, and bioinformatic analysis and three-dimensional modeling predicted their DNA methylation activity. This confirms that I. ricinus possesses a complete DNA methylation toolkit. Our results suggest that DNA methylation is important for the physiology and transstadial development of ticks.
Kateryna Kotsarenko, Valentyna Lylo, Tetiana Ruban, Larysa Macewicz, and Lyubov Lukash
Springer Science and Business Media LLC
The inducible repair enzyme O6-methylguanine-DNA methyltransferase (MGMT) eliminates O6-methylguanine adducts in DNA and protects the cells from damaging effects of alkylating agents. We have found that anti-MGMT antibodies recognize both the MGMT protein with a mol. weight ~ 24 kDa and a protein with a mol. weight ~ 48 kDa, which was named MARP (anti-methyltransferase antibody recognizable protein). A number of growth factors and cytokines were shown to regulate the expression of MGMT and MARP proteins. The ranges of concentrations of several growth factors and cytokines that caused increasing or decreasing protein amounts in human cell cultures were determined. The results of special biological experiments have allowed us to assume a possible role of MARP in the repair of alkyl adducts in human cells.
Kateryna Kotsarenko, Valentyna Lylo, Tetiana Ruban, Larysa Macewicz, and Lyubov Lukash
Springer Science and Business Media LLC
The inducible repair enzyme O6-methylguanine-DNA methyltransferase (MGMT) eliminates O6-methylguanine adducts in DNA and protects the cells from damaging effects of alkylating agents. We have found that anti-MGMT antibodies recognize both the MGMT protein with a mol. weight ~ 24 kDa and a protein with a mol. weight ~ 48 kDa, which was named MARP (anti-methyltransferase antibody recognizable protein). A number of growth factors and cytokines were shown to regulate the expression of MGMT and MARP proteins. The ranges of concentrations of several growth factors and cytokines that caused increasing or decreasing protein amounts in human cell cultures were determined. The results of special biological experiments have allowed us to assume a possible role of MARP in the repair of alkyl adducts in human cells.
Aleksei A. Stepanenko, Svitlana V. Andreieva, Kateryna V. Korets, Dmytro O. Mykytenko, Vladimir P. Baklaushev, Nataliya L. Huleyuk, Oksana A. Kovalova, Kateryna V. Kotsarenko, Vladimir P. Chekhonin, Yegor S. Vassetzky,et al.
Springer Science and Business Media LLC
BackgroundTemozolomide (TMZ) is a first-line drug for the treatment of glioblastoma. Long-term TMZ-treated tumour cells acquire TMZ resistance by profound reprogramming of the transcriptome, proteome, kinome, metabolism, and demonstrate versatile and opposite changes in proliferation, invasion, in vivo growth, and drug cross-resistance. We hypothesized that chromosomal instability (CIN) may be implicated in the generation of TMZ-driven molecular and phenotype diversity. CIN refers to the rate (cell-to-cell variability) with which whole chromosomes or portions of chromosomes are gained or lost.MethodsThe long-term TMZ-treated cell lines were established in vitro (U251TMZ1, U251TMZ2, T98GTMZ and C6TMZ) and in vivo (C6R2TMZ). A glioma model was achieved by the intracerebral stereotactic implantation of C6 cells into the striatum region of rats. Genomic and phenotypic changes were analyzed by conventional cytogenetics, array CGH, trypan blue exclusion assay, soft agar colony formation assay, scratch wound healing assay, transwell invasion assay, quantitative polymerase chain reaction, and Western blotting.ResultsLong-term TMZ treatment increased CIN-mediated genomic diversity in U251TMZ1, U251TMZ2 and T98GTMZ cells but reduced it in C6TMZ and C6R2TMZ cells. U251TMZ1 and U251TMZ2 cell lines, established in parallel with a similar treatment procedure with the only difference in the duration of treatment, underwent individual phenotypic changes. U251TMZ1 had a reduced proliferation and invasion but increased migration, whereas U251TMZ2 had an enhanced proliferation and invasion but no changes in migration. U251TMZ1 and U251TMZ2 cells demonstrated individual patterns in expression/activation of signal transduction proteins (e.g., MDM2, p53, ERK, AKT, and ASK). C6TMZ and C6R2TMZ cells had lower proliferation, colony formation efficiency and migration, whereas T98GTMZ cells had increased colony formation efficiency without any changes in proliferation, migration, and invasion. TMZ-treated lines demonstrated a differential response to a reduction in glucose concentration and an increased resistance to TMZ re-challenge but not temsirolimus (mTOR inhibitor) or U0126 (MEK1/2 inhibitor) treatment.ConclusionLong-term TMZ treatment selected resistant genotype-phenotype variants or generated novel versatile phenotypes by increasing CIN. An increase of resistance to TMZ re-challenge seems to be the only predictable trait intrinsic to all long-term TMZ-treated tumour cells. Changes in genomic diversity may be responsible for heterogeneous phenotypes of TMZ-treated cell lines.
V.V. Lylo, I.S. Karpova, K.V. Kotsarenko, L.L. Macewicz, T.O. Ruban, and L.L. Lukash
International Society for Horticultural Science (ISHS)
Genomes of living organisms are constantly affected by exogenous and endogenous factors, which lead to generating cytotoxic, carcinogenic, and/or mutagenic DNA lesions. However cells possess a number of protective mechanisms directed against DNA damage. The repair enzyme 06-methylguanine-DNA methyltransferase (MGMT) plays a key role in the repair of primary damages of DNA caused by alkylating compounds, which are widely used in industry and medicine. In humans MGMT protects the integrity of the genome, but also contributes to the resistance of tumors to DNA-alkylating chemotherapeutic agents. Therefore, modulation of MGMT expression is a possible strategy to improve the efficiency of cancer therapy and defend normal cells from toxicity of alkylating drugs...
I.S. Karpova, V.V. Lylo, L.L. Macewicz, K.V. Kotsarenko, L.G. Palchykovska, T.O. Ruban, and L.L. Lukash
International Society for Horticultural Science (ISHS)
The human genome is affected by diverse environmental pollution factors and this is particularly acute for Ukrainians who have suffered from the Chernobyl radiation disaster. At the same time many natural compounds, especially from medicinal plants, are known as agents that prevent DNA lesions which often result in increasing mutagenesis and carcinogenesis. The aim of our work was to study the DNA-protective potential of biologically active compounds found in Sambucus nigra called lectins, a very large group of universally occurring proteins that recognize and specifically bind to carbohydrates/glycoconjugates. Various physiological activities of lectins based on protein-carbohydrate interactions have been demonstrated in all living forms. For example, lectins participate in host defense in plants against stress-related conditions, the attack of phytopathogens and phytophagous insects, as well as modulation of immune response, mitogenic stimulation or induction of apoptosis in animals. Antiviral, immunomodulating, antioxidant and insulin-stimulating properties of S. nigra fruit and flower extracts have been described in scientific literature. Also the elderberry lectins were found in roots, leaves, bark, seeds and fruits, the latter (SNA-IV) being the predominant protein in the juice. In Ukraine, various parts of S. nigra plants are used in folk medicine. Using methods of isoelectric focusing and chromatography, we determined that elderberry flowers and pollen contain rather high levels of lectins agglutinating animal and human erythrocytes, which differ from a commercial preparation of S. nigra bark lectin (SNA-I). The major lectin found in inflorescences, named SNAflu-I, is GalNAc/Gal specific and is supposed to be a heterotetramer with subunits of about 30 and 33 kDa. Sambucus nigra lectins demonstrated protective effects against heavy metals (nickel ions) in soil bacteria Bacillus subtilis. Also it was shown that lectins under study can modulate, in a concentration-dependent manner, the frequency of mutations and genotoxic activity of alkylating agents in eukaryotic cell cultures. Studies focused on the elucidation of cell targets sensitive to S. nigra lectins and demonstrated that at least one of the targets may be the DNA-dependent synthesis of RNA â the way to modulate gene expression. The results obtained lead to the conlusion that the protective functions of lectins both in pro- and eukaryotes involve complex mechanisms including components of the DNA repair system.
K. V. Kotsarenko, V. V. Lylo, T. P. Ruban, L. L. Macewicz, A. I. Kornelyuk, S. I. Chernykh, and L. L. Lukash
Institute of Molecular Biology and Genetics (NAS Ukraine)
Aim. To study the effect of EMAP II, IFN2b and its medicinal preparations on the amount of O 6 -methylguanine-DNA methyltransferase (MGMT) protein in human cells in vitro. Methods. The human cells of 4BL and Hep-2 lines were treated with the purified recombinant proteins EMAP II, IFN2b and its commercial me dicinal preparations. Changes in the MGMT gene expression were studied at a protein level by Western blot analysis. Results. Treatment of Hep-2 and 4BL cells with EMAP II at the concentrations of 0.02 g/ml and 2 g/ml respectively led to induction of the MGMT gene expression. EMAP II at the concentrations of 0.2–20 g/ml caused decrease of the MGMT protein amount in Hep-2 cells. The regulating activity of EMAP II was also observed for MARP (anti-Methyltransferase Antibody Recognizable Protein). IFN2b and Laferon-PharmBiotek with the activity of 200 and 2000 IU/ml were shown to cause an increase of the MGMT protein amount in Hep-2 cells. Conclusions. The purified recombinant proteins EMAP II and IFN2b which are substrates for the medicinal preparations influenced on the amount of MGMT protein in the human cell cultures in a concentration-dependent manner. At the same time the effect of medicinal preparations differs from that of the purified protein IFN2b. Possibly it depends on the presence of stabilizing components in their compositions.
K. V. Kotsarenko, V. V. Lylo, L. L. Macewicz, T. P. Ruban, Yu. S. Luchakivska, M. V. Kuchuk, and L. L. Lukash
Institute of Molecular Biology and Genetics (NAS Ukraine)
Aim. To investigate an effect of biologically active compounds IFN2b, EMAPII, Card medium, fibronectin on the amount of MGMT (O-methylguanine-DNA methyltransferase) and MARP (anti-Methyltransferase Antibody Recognizable Protein) proteins in human cells in vitro. Methods. The human cells of 4BL, Hep-2 and A102 lines were treated with growth factors and cytokines. Changes in the amount of MGMT and MARP proteins were studied by Western blot analysis with anti-MGMT mAbs. Results. The treatment of A102 cells with EMAPII, fibronectin, Laferon and Card medium led to a decreased level of the MGMT protein, whereas the amount of MARP was highly increased in these cells. The treatment with the recombinant protein IFN2b increased the amount of MGMT and MARP proteins in Hep-2 cells. The treatment with extracts of transgenic plants,containing human IFN2b, caused a significant decrease in the content of both proteins in Hep-2 cells and MARP in 4BL cells. Conclusions. Both MGMT and MARP are highly inducible proteins. Their amount in cells can be changed by some growth factors (Card medium, fibronectin), cytokine (IFN2b), cytokine-like (EMAPII) or cytokine-containing substances (Laferon and IFN2b in plant extracts). This regulation depended not only on the type of biologically active substances but on the cell line used in this study as well.
L. L. Macewicz, V. O. Kushniruk, A. P. Iatsyshyna, K. V. Kotsarenko, V. V. Lylo, G. R. Akopyan, N. L. Huleuk, D. O. Mykytenko, and L. L. Lukash
Institute of Molecular Biology and Genetics (NAS Ukraine)
Long-term cultivation of cell lines inevitably leads to genetic and epigenetic changes. Aim. A comparative analysis of karyotype and level of the expression of reparative enzyme O 6 -methylguanine-DNA-methyltransferase (MGMT) at different stages of establishment and stabilization of human cell line 4BL and cell line of mouse germ cells G1. Methods. The set of methods was used to research the dynamics of karyotypes changes: the differential staining of chromosomes, FISH-method and comparative genomic hybridization. The level of MGMT expression was analyzed by PCR reaction and Western blot analysis. Results. General trends of establishment of mouse and human cell lines were revealed: at the first stage, which is characterized by increased structural instability of the genome, an increase in the MGMT expression was revealed while at the second stage of stabilization – a decrease in the expression. Therefore, almost complete disappearance of MGMT protein in unmodified form (24 kDa) is observed. Conclusions. Statistically significant correlation between MGMT repair enzyme and mutations induction processes during mammalian cell adaptation and cell line establishment to in vitro was described.
K. V. Kotsarenko, V. V. Lylo, L. L. Macewicz, L. A. Babenko, A. I. Kornelyuk, T. A. Ruban, and L. L. Lukash
Allerton Press
The influence of cytokines LIF, SCF, IL-3, and EMAP II and the Laferobion (IFN-a2b) drug on the MGMT gene expression in human cell cultures has been studied. It was shown that exogenous cytokines can modulate the MGMT gene expression at the protein level. EMAP II is able to increase or decrease the MGMT level, depending on the experimental conditions. Cytokines LIF, SCF, IL-3 and Laferobion decreased the MGMT expression level in human cells in vitro. Some conditions leading to the destruction of the MGMT protein complex were identified.
V. V. Lylo, L. L. Matsevich, E. V. Kotsarenko, L. A. Babenko, A. I. Kornelyuk, E. M. Sukhorada, and L. L. Lukash
Allerton Press
The aim of the study is to evaluate the effect of recombinant EMAP II cytokine (endothelial and monocyte-activating polypeptide II) on the level of MGMT gene expression; this gene encodes the O6-methylguanine-DNA-methyltransferase (MGMT) repair enzyme in the cell culture of humans. An investigation into the EMAP II effect on the proliferation of cells was carried out using the standard MTT test. The MGMT protein in a cell extract was identified by Western blot analysis. The following cell lines were investigated: A102 (fibroblasts), CB-1 (umbilical cord blood stromal cells), and 4BL6 (cells obtained from peripheral blood). It was shown in these experiments that the EMAP II cytokine induces MGMT expression in human cells of the investigated lines. There was observed a decrease in the quantity of cells in the presence of a high concentration of this cytokine. The level of expression of the MGMT repair enzyme was established to increase in human cells in vitro in a serum-free culture medium with the EMAP II cytokine.