Papers by Serhiy Souchelnytskyi
Acta Biochimica Polonica, Jun 30, 2003
Acquired drug resistance of tumor cells is frequently observed in cancer patients undergoing chem... more Acquired drug resistance of tumor cells is frequently observed in cancer patients undergoing chemotherapy. We studied murine leukemia L1210 cells sensitive and resistant to the cytotoxic action of cisplatin and showed that cisplatin-resistant leukemia cells were also refractory to TGF b1-dependent growth inhibition and apoptosis. Addressing the question about the mechanisms responsible for the cross-resistance to cisplatin and TGF b1, we found that cisplatin-and TGF b1-resistant L1210 cells possessed a decreased expression of type I TGF b1 receptor, while the expression of type II TGF b1 receptor was not affected. Western blot analysis of Smad proteins 2, 3, 4, 6, and 7, which participate in signal transduction pathway downstream of the TGF b1 receptors, revealed an increased expression of Smad 6, inhibiting TGF b1 action, only in cisplatin-and TGF b1-resistant L1210 cells. TGF b1 and especially the cytotoxic mistletoe agglutinin increased Smad 6 expression in TGF b1-sensitive but not in TGF b1-resistant L1210 cells. TGF b1-resistant L1210 cells also differed from TGF b1-sensitive cells by the lack of expression of the pro-apoptotic p53 protein and higher
Pathobiology, 2018
The aim of this study was to identify differences in proteome profiles of diffuse large B-cell ly... more The aim of this study was to identify differences in proteome profiles of diffuse large B-cell lymphoma (DLBCL) of nongerminal center (non-GC) versus GC type in the search for new markers and drug targets. Methods: Six DLBCL, with 3 repeats for each, were used for the initial study by proteomics: 3 non-GC and 3 GC DLBCL cases. For immunohistochemistry, tissue microarrays were made from 31 DLBCL samples: 16 non-GC de novo lymphomas and 15 GC cases (11 transformed from follicular lymphomas and 4 de novo GC lymphomas). Proteome profiling was performed by two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry. Results: Ninety-one proteins were found differentially expressed in non-GC compared to GC type. The Cytoscape tool was used for systemic analysis of proteomics data, revealing 19 subnetworks representing functions affected in non-GC versus GC types of DLBCL. Conclusion: A validation study of 3 selected proteins (BiP/Grp78, Hsp90, and cyclin B2) showed the enhanced expression in non-GC DLBCL, supporting the proteomics data.
Ukrainian Biochemical Journal, Oct 4, 2022
Blood sera of 12 severe covid-19 patients and 14 healthy human donors were subjected to original ... more Blood sera of 12 severe covid-19 patients and 14 healthy human donors were subjected to original tca-extraction/acetone-precipitation followed by SDS-PaaG electrophoresis and mass-spectrometry. 76 kDa protein was detected as one of the differentially expressed proteins in the samples of Covid-19 patients. This 76 kDa protein was identified with mass-spectrometry as human serum albumin. Such molecular form of albumin was absent in blood serum of healthy human donors. the potential ways of generation of the unusual form of human serum albumin and its probable diagnostic value were discussed.
Journal of Proteomics & Bioinformatics, Nov 25, 2016
Journal of Proteomics & Bioinformatics, Nov 25, 2016
Праці Наукового товариства імені Шевченка, Nov 23, 2020
Background. COVID-19 pandemic highlighted the importance of sensitive and specifi c tests that wo... more Background. COVID-19 pandemic highlighted the importance of sensitive and specifi c tests that would be cost-effi cient, fast and scalable. There are more than 200 COVID-19 detection tests available worldwide, with every country developing its own assays. Sample collection, preparing for a test, the test itself and interpretation of results have a strong impact on the clinical value of testing. The diversity of tests and workfl ows requires the analysis of their performance in clinics. Methods. Literature review, analysis of clinical reports, online resources, public and commercial reports were used to collect information about tests. The collected information was processed to obtain information relevant to this review. Results. COVID-19 tests based on the amplifi cation of nucleic acids are reviewed. Tests employ polymerase chain reaction (PCR) or loop-mediated isothermal amplifi cation (LAMP). The clinical value of these tests depends on the technologies used, as they diff er for LAMP, real-time and standard PCR methods. The diversity of sample preparation protocols, diff erent designs of the tests, used chemicals and protocols have a significant impact on tests. Tailoring a testing workfl ow to available infrastructure and selecting the most effi cient combination of tests and protocols for each step in a testing workfl ow is crucial for the success. Conclusion. Strengths and weaknesses of diff erent test systems and protocols that were reviewed herein can be helpful in selecting a testing workfl ow to achieve maximum clinical utility.
Pracì Naukovogo tovaristva ìmeni Ševčenka, Apr 15, 2020
Diagnostic based on analysis of living tumor cells is frequently used in oncology. Circulating tu... more Diagnostic based on analysis of living tumor cells is frequently used in oncology. Circulating tumor cells in the blood and cells obtained from a tumor biopsy are used to access their carcinogenic properties and subsequently to predict possible development of the disease. Here we report use of these two tests to assess aggressiveness and metastatic potential of a bronchial adenocarcinoma. The circulating tumor cells test was negative, no circulating cells was observed. It indicates that there was no metastatic spread. However, test with the surgery biopsy showed presence of aggressive cellular clones. The tumor cells from the biopsy proliferated and spread from the cultured tissue. Moreover, the tumor cells formed colonies of cells which lost contact inhibition. This is an indication of aggressive carcinogenic features of the cells in tumor organoids. Combination of both tests showed that the local tumor had an aggressive phenotype, but no detectable spreading of cells. Therefore, these tests support a management plan with removal the primary tumor and regular monitoring, without need of an extensive chemotherapy.
Current Protein & Peptide Science, Feb 17, 2022
: Post-translational modifications (PTMs) may affect functions of human serum albumin. Here we re... more : Post-translational modifications (PTMs) may affect functions of human serum albumin. Here we review reports of novel PTMs of human serum albumin. One hundred twenty-three recently reported novel O-phosphorylation, glycation, methylation, carbonylation, and acetylation of albumin are reviewed. Potential impact of these PTMs on albumin functions is discussed. Knowledge of these PTMs of albumin is of importance for use of albumin in medical applications, e.g., in transfusion, drug formulations and remedies.
Research Square (Research Square), Sep 1, 2020
BACKGROUND Chloroquine is used for the treatment of COVID-19 patients. However, e cacy of the chl... more BACKGROUND Chloroquine is used for the treatment of COVID-19 patients. However, e cacy of the chloroquine has been under discussion. Variability of clinical outputs of the drug application requires implementation of a companion diagnostic that would allow monitoring responsiveness to chloroquine. The rst line of such markers would be markers already used in clinics. Analysis of reported mechanisms of COVID-19 and chloroquine may lead to such markers. METHODS Systemic analysis of molecular mechanisms and markers engaged by chloroquine and COVID-19 virus was performed. The networks of regulatory mechanisms were explored for an intersection and relevance to clinical markers. RESULTS Reported here systemic analysis describes the intersection of molecular mechanisms of chloroquine and processes engaged by COVID-19. 266 nodes provide insight into the mechanisms of chloroquine impact on the infection and represent a pool of companion diagnostic markers. As an example, an intersection with the markers of heart arrhythmia retrieved 19 nodes. Thirteen of them were reported in human plasma: levels of albumin, amyloid precursor protein, and endoglin correlate with adverse cardiac effects. CONCLUSIONS Reported nodes are the candidate markers for companion diagnostic of the chloroquine application to COVID-19 patients. Some of these markers are already used in the clinic and their interpretation may contribute to monitoring for adverse effects of chloroquine.
Reproductive Biology, Mar 1, 2021
The dynamic embryo development during the early stages of gestation requires precise molecular ch... more The dynamic embryo development during the early stages of gestation requires precise molecular changes, including proteomic ones. We aimed to find unique proteins for porcine conceptuses specifically during the peri-implantation period, i.e. on days 15-16 of pregnancy. The proteomic profile of these conceptuses was compared with conceptuses at an earlier stage of the development, i.e. collected during maternal recognition of pregnancy on days 12-13 of pregnancy. The 2DE, gel image analysis, and MALDI TOF mass spectrometry were used 500 protein spots were annotated as common to conceptuses harvested during both studied periods. Proteomic profile of the conceptuses collected during the peri-implantation period contains 24 unique proteins. Identified unique for the peri-implantation period proteins are involved in adhesion processes, cadherin, and actin-binding, and actin filament organization, extracellular matrix organization, and cytoskeleton organization. Systemic analysis of identified proteins confirmed their involvement in cell adhesion and cytoskeletal organization as being two major affected functions. The unique proteins might be recognized as factors conditioning the proper peri-implantation embryo development and gaining competences for implantation. In further studies, BRCA1 might be considered as a candidate for a potential marker of embryonic competences for implantation in pigs.
Experimental Oncology, 2019
Transforming growth factor β1 (TGF β1) is a potent regulator of breast tumorigenesis. It inhibits... more Transforming growth factor β1 (TGF β1) is a potent regulator of breast tumorigenesis. It inhibits proliferation of carcinoma cells, but the strength of its inhibitory action varies for cells from benigh, non-metastatic or metastatic tumors. The aim of this work was to generate a proteome profile of TGF β1 action on non-tumorigenic human breast epithelial cells 184A1, and validate predicted involvement of casein kinase 2α (CK2α), p53 and structure-specific recognition protein-1 (SSRP1). Materials and Methods: Two-dimensional gel electrophoresis and mass spectrometry were used to identify TGF β1-regulated proteins in 184A1 human breast immortalized non-tumorigenic cells. 184A1 cells may serve as a model of benign breast neoplasia. These cells were obtained from normal mammary tissue, were immortalized but are not malignant, and were obtained from the American Type Culture Collection. The systemic analysis was performed by using the Cytoscape tool. Transfection of cells with CK2α construct and small interfering RNAs to CK2α and SSRP1 were used to assess an impact of CK2α and SSRP1 on phosphorylation of the p53 and cell proliferation. Results: Proliferation of 184A1 cells was transiently inhibited by TGF β1. We identified 100 and 47 unique proteins which changed their expression and/or 35 S-incorporation, respectively, upon treatment with TGF β1 for 2 h, 8 h or 24 h. Cell proliferation, death, migration, and metabolism were among the biological regulatory processes retrieved by the network analysis as affected by the identified proteins. The network analysis suggested that TGF β1 may affect the phosphorylation of p53 at Ser392 by engaging CK2α. This was confirmed by the immunoblotting and cell proliferation assays. Conclusion: We report here the list of 147 TGF β1-regulated proteins in immortalized non-tumorigenic human breast epithelial cells, and show involvement of CK2α in the regulation of p53 Ser392 phosphorylation.
OncoTargets and Therapy, Aug 1, 2014
The response of cells to TGFβ and EGF is mediated by a network of various intracellular regulator... more The response of cells to TGFβ and EGF is mediated by a network of various intracellular regulators. The signaling crosstalk between different regulators is of key importance for tumorigenesis. The crosstalk may explain the modulation of cellular responses to the same regulator by another signaling molecule. As PKN1-a serine/threonine kinase implicated in tumorigenesis-was identified as potential crosstalk node for TGFβ and EGF signaling, the cellular functions that may be affected by PKN1 in a crosstalk of TGFβ and EGF were explored. Methods: To investigate the contribution of PKN1 to TGFβ and EGF signaling, transiently PKN1-transfected HEC-1-A endometrial cancer cells were generated and subjected to treatment with TGFβ1, EGF, and their combination. Proliferation, apoptosis, invasion, wound healing, and migration assays were performed. The impact of PKN1 on the expression and phosphorylation of intracellular proteins was monitored by immunoblotting. Results: It was demonstrated that PKN1 modulated the responses of HEC-A-1 endometrial cancer cells to TGFβ1 and EGF. PKN1 had an inhibitory effect on the stimulation of cell migration, and PKN1 kinase activity was required for the inhibitory effect of TGFβ and EGF on cell proliferation and invasiveness. It was observed that phosphorylation of Smad2, FAK, and Erk1/2 correlated with responses of the cells to TGFβ1 and EGF. Conclusion: PKN1 modulates TGFβ-and EGF-dependent regulation of cell proliferation, migration, and invasiveness, and therefore is a component of the network signaling downstream of TGFβ and EGF.
PubMed, Dec 1, 2022
Purpose: Serum albumin is in contact with practically all cells in the human body, including tumo... more Purpose: Serum albumin is in contact with practically all cells in the human body, including tumor cells in cancer patients. The purpose of this study was to explore whether cancer cells affect post-translational modifications (PTMs) of albumin. Material and methods: Mass spectrometry was used to identify the PTMs. Purified human serum albumin was incubated with human breast cancer cells MDA-MB-231, MDA-MB-468, MCF7, or kept in water or in cell culture media. PTMs which were affected upon exposure of the albumin to cancer cells were identified. Three-dimensional analysis was performed to locate PTMs in albumin. Results: We report here that an exposure to human breast cancer cells affected post-translational modifications (PTMs) of 14 peptides of human serum albumin (HSA). PTMs at 8 peptides were observed upon exposure of HSA to metastatic MDA-MB-231 and MDA-MB-468 breast cancer cells. PTMs at another 6 peptides were lost in MDA-MB-231 and MDA-MB-468 cells, while these 6 PTMs were observed in HSA exposed to conditionally tumorigenic MCF7 cells, or in HSA kept in water or a cell culture medium. Cancer cell altered phosphorylation, deamidation followed by methylation, acetylation, myristylation, palmitoylation, methylation, cysteine persulfide, and S-6-FMN cysteine modifications were detected in HSA. These PTMs locate predominantly in IB and IIA domains of HSA. Three-dimensional analysis showed that this region corresponds to the lipid-binding site and Sudlow's site 1. Conclusion: Data reported here show that 14 PTMs of human serum albumin can be modified upon its exposure to human breast cancer cells.
Experimental Oncology, 2002
Transforming growth factor-b (TGFb) is a polypeptide growth factor, which regulates cell prolifer... more Transforming growth factor-b (TGFb) is a polypeptide growth factor, which regulates cell proliferation, differentiation, migration and apoptosis. During the last decade an exploration of mechanisms ...
Journal of Elementology, Jul 11, 2016
<p>(<b>A</b>) Graphs show distribution of connections for species of the networ... more <p>(<b>A</b>) Graphs show distribution of connections for species of the network. Distribution for proteins identified by phosphoproteomics (rhombs; experimental data) and as would be expected by a power law distribution of connections of species in an ideal scale-free network (squares; predicted distribution) are shown. (<b>B</b>) A sub-network of 14-3-3σ (SFN). Proteins which are in proximal dependencies to 14-3-3σ were extracted from the complete network (<a href="https://www.plosone.org/article/info:doi/10.1371/journal.pone.0065163#pone.0065163.s005" target="_blank">Figure S5</a>) into the presented sub-network. (<b>C</b>) Signaling pathways involving the proteins assembled in the 14-3-3σ sub-network. The analysis was performed using Gene Set Analysis Toolkit V2 software. (<b>D</b>) Validation of 14-3-3σ phosphorylation upon treatment of cells with TGFβ1. Images of the areas of 2D Fe-IMAC gels with annotation of phosphoprotein spots p72 and p74, in which 14-3-3σ was identified are shown. Values of the protein spot volumes are shown below images of gels for both protein spots. (<b>E</b>) Phosphorylation of endogenous 14-3-3σ in MCF10A cells was evaluated by immunoprecipitation with anti-phosphoserine, threonine and tyrosine antibodies (upper panel) or by incorporation of <sup>32</sup>P (middle panel). Control immunoprecipitation of 14-3-3σ is shown in lower panel. Densitometry analysis of the protein immunoblots or <sup>32</sup>P incorporation is shown in accompanying graphs. (<b>F</b>) Phosphorylation of Flag-14-3-3σ fusion protein expressed in HEK293T cells was evaluated in the same way as for endogenous protein. The upper panel shows detection of phosphorylation by immunoprecipitation and the middle panel shows incorporation of <sup>32</sup>P. The lower panel shows expression of 14-3-3σ. Migration positions of 14-3-3σ are shown by the arrows and treatments with TGFβ1 are indicated. Densitometry analysis of the protein immunoblots is shown in accompanying graphs. Representative experiments out of 3 performed are shown.</p
Biochem Biophys Res Commun, 2002
Experimental oncology
Aim: to study in vitro the antiproliferative influence of interferon-α2b (²FN-α2b), transforming ... more Aim: to study in vitro the antiproliferative influence of interferon-α2b (²FN-α2b), transforming growth factor-β1 (TGF-β1), cys-platinum, cyclophosphamide, and paclitaxel on the of human microvascular endothelial cells for the development of approaches of the antiangiogenic therapy of malignancies. Methods: [3H]-methylthymidine incorporation into human microvascular endothelial derma-derived cells (HMVECd) treated with ²FN-a2b, TGF-b1, cysplatinum, cyclophosphamide, and paclitaxel have been studied. Results: studied cytokines and drugs inhibited proliferation of HMVECd cells at low concentrations (²FN-α2b — 50.0 and 100.0 IU/ml; TGF-β1 — 5.0 ng/ml; cysplatinum — 0.5 μg/ml; paclitaxel — 10.0 μg/ml; cyclophosphamide — 50.0 μg/ml). Conclusion: low concentrations of ²FN-α2b, ÒGF-β1, cysplatinum, cyclophosphamide, ànd paclitaxel inhibited proliferation of cultured endothelial cells, suggesting a possibility of low-dose antiangiogenic therapy in the long-term (“metronomic”) regimen.
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Papers by Serhiy Souchelnytskyi