Publications by authors named "Tomasz Kubrak"

10 Publications

  • Page 1 of 1

The Correlation of Mutations and Expressions of Genes within the PI3K/Akt/mTOR Pathway in Breast Cancer-A Preliminary Study.

Int J Mol Sci 2021 Feb 19;22(4). Epub 2021 Feb 19.

Chair and Department of Biology and Genetics, Medical University of Lublin, 20-093 Lublin, Poland.

There is an urgent need to seek new molecular biomarkers helpful in diagnosing and treating breast cancer. In this elaboration, we performed a molecular analysis of mutations and expression of genes within the PI3K/Akt/mTOR pathway in patients with ductal breast cancer of various malignancy levels. We recognized significant correlations between the expression levels of the studied genes. We also performed a bioinformatics analysis of the data available on the international database TCGA and compared them with our own research. Studies on mutations and expression of genes were conducted using High-Resolution Melt PCR (HRM-PCR), Allele-Specific-quantitative PCR (ASP-qPCR), Real-Time PCR molecular methods in a group of women with ductal breast cancer. Bioinformatics analysis was carried out using web source Ualcan and bc-GenExMiner. In the studied group of women, it was observed that the prevalence of mutations in the studied and genes was 29.63%. It was stated that the average expression level of the , , genes in the group of breast cancer patients is lower in comparison to the control group, while the average expression level of the and genes in the studied group was higher in comparison to the control group. It was also indicated that in the group of patients with mutations in the area of the and genes, the gene expression level is statistically significantly lower than in the group without mutations. According to our knowledge, we demonstrate, for the first time, that there is a very strong positive correlation between the levels of and gene expression in the case of patients with mutations and without mutations.
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http://dx.doi.org/10.3390/ijms22042061DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7922286PMC
February 2021

Imaging Flow Cytometric Analysis of Stilbene-Dependent Apoptosis in Drug Resistant Human Leukemic Cell Lines.

Molecules 2019 May 17;24(10). Epub 2019 May 17.

Clinical Genetics Department, Medical University of Lublin, 11 Radziwillowska Str., 20-080 Lublin, Poland.

The natural compounds have been researched extensively as an alternative to the conventional chemotherapy and radiation. Stilbene derivatives appear as a group of therapeutics which deserves special attention. The present study was designed to analyze the effects of stilbene derivatives on drug resistant human leukemic cells. The aim of this work was to evaluate the apoptotic effect of stilbene derivatives in various concentrations on leukemic cells (LC) with and without resistant phenotype. Human acute promyelocytic leukemia (APL) cell lines (HL60, HL60/MX1, HL60/MX2) and acute lymphoblastic leukemia (ALL) cell lines (CEM/C1, CCRF-CEM) were studied. T-resveratrol, piceatannol, rhaponticin, deoxyrhaponticin, pterostilbene were used to stimulate apoptosis. Mitoxantrone (MIT) was applied to induce drug resistance. -Resveratrol (RES), deoxyrhaponticin (D-RHAP), rhaponticin (RHAP), pterostilbene (PTER), and piceatannol (PIC) influenced viability and induced apoptosis in all investigated cell lines. Our results confirmed that RES, PIC, RHAP, D-RHAP, and PTER are essential therapeutic compounds with anticancer activity exhibited by induction of apoptosis in leukemic cells with and without resistant phenotype. Stilbene-induced apoptosis in HL60/MX1, HL60/MX2, CEM/C1, and CCRF-CEM leukemia cell lines have been presented in very few studies so far and our research is an important contribution to the investigation of these substances.
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http://dx.doi.org/10.3390/molecules24101896DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6571880PMC
May 2019

The Effect of Furanocoumarin Derivatives on Induction of Apoptosis and Multidrug Resistance in Human Leukemic Cells.

Molecules 2019 May 12;24(9). Epub 2019 May 12.

Chair and Department of Biology and Genetics, Faculty of Pharmacy with Medical Analytics Division, Medical University of Lublin, 20-059 Lublin, Poland.

Background: The insensitivity of cancer cells to therapeutic agents is considered to be the main cause of failure of therapy and mortality of patients with cancer. A particularly important problem in these patients is the phenomenon of multidrug resistance, consisting of abnormal, elevated expression of transport proteins (ABC family). The aim of this research included determination of IC50 values of selected furanocoumarins in the presence and absence of mitoxantrone in leukemia cells and analysis of changes in apoptosis using anexinV/IP and Casp3/IP after 24 h exposure of cell lines to selected coumarins in the presence and absence of mitoxantrone in IC50 concentrations.

Methods: Research was conducted on 3 cell lines derived from the human hematopoietic system: HL-60, HL-60/MX1 and HL-60/MX2. After exposure to coumarin compounds, cells were subjected to cytometric analysis to determine the induction of apoptosis by two methods: the Annexin V test with propidium iodide and the PhiPhiLux-G1D2 reagent containing caspase 3 antibodies.

Results: All of the furanocoumarin derivatives studied were found to induce apoptosis in leukemia cell lines.

Conclusions: Our results clearly show that the furanocoumarin derivatives are therapeutic substances with antitumor activity inducing apoptosis in human leukemia cells with phenotypes of resistance.
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http://dx.doi.org/10.3390/molecules24091824DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6539621PMC
May 2019

Separation and determination of coumarins including furanocoumarins using micellar electrokinetic capillary chromatography.

Talanta 2018 Sep 8;187:120-124. Epub 2018 May 8.

Department of Analytical Chemistry, Medical University of Lublin, Chodźki 4a, 20-093 Lublin, Poland.

The conditions of micellar electrokinetic capillary chromatography for separation and simultaneous measurement of coumarins (coumarin, scoparone, isoscopoletin, esculin, esculetin, umbelliferone) including furanocoumarins (xanthotoxin, byakangelicin, isopimpinellin, bergapten, phellopterin, xanthotoxol) have been elaborated. The influence of different parameters, such as the pH of the buffer, sodium cholate (SC) or methanol concentration in the buffer, on the migration time, peak resolution, peak asymmetry, and number of theoretical plates was investigated. The optimum separation of the compounds was achieved using 50-µm i.d. capillaries with a total length of 64.5 cm (56 cm effective length) and a buffer system at pH 9.00 consisting of 50 mM sodium tetraborate, 45 mM SC, and 20% of methanol (v/v). The developed method ensured good repeatability of corrected peak areas and migration times (the relative standard deviations were in the range of 2.8-6.1% and 0.8-4.0%, respectively). The average limit of detection for all studied compounds was below 1.3 µg mL. Moreover, good linearity of the relationship between the peak corrected area and the concentration of the compounds was observed (correlation coefficient >0.99). The method was successfully applied in the quantitative analysis of two different types of samples, i.e. Heracleum sphondylium herb and Aesculus hippocastanum cortex.
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http://dx.doi.org/10.1016/j.talanta.2018.05.024DOI Listing
September 2018

Modulation of Multidrug Resistance Gene Expression by Coumarin Derivatives in Human Leukemic Cells.

Oxid Med Cell Longev 2017 13;2017:5647281. Epub 2017 Dec 13.

Department of Clinical Genetics, Medical University of Lublin, Lublin, Poland.

The presence of multidrug resistance (MDR) in tumor cells is considered as the major cause of failure of cancer chemotherapy. The mechanism responsible for the phenomenon of multidrug resistance is explained, among others, as overexpression of membrane transporters primarily from the ABC family which actively remove cytostatics from the tumor cell. The effect of 20 coumarin derivatives on the cytotoxicity and expression of , , , and genes (encoding proteins responsible for multidrug resistance) in cancer cells was analyzed in the study. The aim of this research included determination of IC10 and IC50 values of selected coumarin derivatives in the presence and absence of mitoxantrone in leukemia cells and analysis of changes in the expression of genes involved in multidrug resistance: , , , and after 24-hour exposure of the investigated cell lines to selected coumarins in the presence and absence of mitoxantrone in IC10 and IC50 concentrations. The designed research was conducted on 5 cell lines derived from the human hematopoietic system: CCRF/CEM, CEM/C1, HL-60, HL-60/MX1, and HL-60/MX2. Cell lines CEM/C1, HL-60/MX1, and HL-60/MX2 exhibit a multidrug resistance phenotype.
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http://dx.doi.org/10.1155/2017/5647281DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5745744PMC
August 2018

Selected secondary metabolites in Echium vulgare L. populations from nonmetalliferous and metalliferous areas.

Phytochemistry 2017 Jan 14;133:4-14. Epub 2016 Nov 14.

Department of Plant Physiology, Maria Curie-Skłodowska University, Akademicka 19, 20-033 Lublin, Poland.

The aim of this study was to evaluate the effect of severe environmental conditions prevailing on metalliferous waste heaps and heavy metal-contaminated growth substrates on accumulation of selected secondary metabolites, antioxidant capacity, and heavy metal concentration in two metallicolous (MC, MZ) and one nonmetallicolous (NM) populations of Echium vulgare L. The shoots and the roots of the three studied populations were collected from their natural habitats. Additionally, the plants were cultivated on different growth substrates, i.e. a contaminated substrate obtained from the areas of growth of the MZ and MC populations and an uncontaminated one from the NM population site. Several compounds, i.e. allantoin, rutin, rosmarinic acid, chlorogenic acid, and 4-hydroxybenzoic acid were identified in the shoots. Moreover, rosmarinic acid, allantoin, and shikonin were measured in the roots. The adverse environmental conditions contributed to a ca. 10- and 4-fold increase in the concentration of allantoin in the roots and shoots, respectively, as well as a ca. 4-fold and ca. 3-fold increase in the level of 4-hydroxybenzoic acid and shikonin, respectively, in comparison with the plants from the uncontaminated site. Similarly, a great impact of the contaminated substrate on the compounds was demonstrated in the soil experiment. Regardless of the populations, even ca. 20-fold higher levels of allantoin and shikonin were observed in plants grown on the MC and MZ substrates. In contrast, the chlorogenic acid concentration was lower in plants collected from the metalliferous areas and in all populations cultivated on the contaminated substrates in comparison with plants from the uncontaminated soil. Unambiguous results were obtained in the case of rutin, i.e. decreased accumulation in both metallicolous populations from the natural environment and increased accumulation in plants grown on the contaminated substrates. The high concentrations of heavy metals in the substrates contributed to high HM concentrations in plant tissues. However, some differences were observed between the metallicolous and nonmetallicolous populations - the accumulation of metals was lower in the shoots and higher in the roots of the NM population, compared with the MZ and MC populations.
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http://dx.doi.org/10.1016/j.phytochem.2016.11.001DOI Listing
January 2017

Comparison of Analytical Methods in Chemometric Fingerprinting of Metallicolous and Non-metallicolous Populations of Echium vulgare L.

Phytochem Anal 2016 Sep 20;27(5):239-48. Epub 2016 Jul 20.

Department of Plant Physiology, Maria Curie-Skłodowska University, Akademicka 19, 20-033, Lublin, Poland.

Introduction: Adverse environmental conditions usually change plant biochemical pathways resulting in accumulation or decreased content of both primary and secondary metabolites. The chemometric fingerprinting analysis proves to be a useful tool to reveal phytochemical differentiation between plants inhabiting heavy metal-contaminated and uncontaminated areas.

Objective: Development and assessment of four analytical techniques - high performance capillary electrophoresis (HPCE), thin-layer chromatography (TLC), mass spectrometry (MS), and Fourier transform infrared (FTIR) spectroscopy in chemometric fingerprinting of metallicolous and non-metallicolous populations of Echium vulgare L.

Material And Methods: Twenty-one crude methanol extracts of shoot samples representing three populations of Echium vulgare L., two originating from highly metal polluted areas and one from an unpolluted area, were investigated using four analytical methods: HPCE, TLC, MS, and FTIR spectroscopy. Data pre-processing (denoising, background subtracting, horizontal alignment) followed by principal component analysis (PCA), hierarchical clustering analysis (HCA), and phytochemical difference index (DI) calculations facilitated exploration of the differences and similarities between the populations.

Results: Clear phytochemical divergence between metallicolous and non-metallicolous populations of Echium vulgare was found. The suitability of the analytical techniques for revealing phytochemical markers and discrimination of individuals originating from different populations differed and in general increased in the order: TLC < MS = HPCE < FTIR.

Conclusion: The chemometric methods applied were successful in discrimination between samples from polluted and unpolluted areas, showing a potential perspective for environmental quality control. Copyright © 2016 John Wiley & Sons, Ltd.
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http://dx.doi.org/10.1002/pca.2620DOI Listing
September 2016

Study of cytotoxic activity, podophyllotoxin, and deoxypodophyllotoxin content in selected Juniperus species cultivated in Poland.

Pharm Biol 2015 Jun 27;53(6):831-7. Epub 2015 Feb 27.

Department of Clinical Genetics, Faculty of Medicine, Medical University of Lublin , Lublin , Poland .

Context: The demand for podophyllotoxin and deoxypodophyllotoxin is still increasing and commercially exploitable sources are few and one of them, Podophyllum hexandrum Royle (Berberidaceae), is a "critically endangered" species.

Objective: The first aim was to quantify the amount of podophyllotoxin and deoxypodophyllotoxin in 61 Juniperus (Cupressaceae) samples. Cytotoxic activity of podophyllotoxin and ethanolic leaf extracts of Juniperus scopulorum Sarg. "Blue Pacific" and Juniperus communis L. "Depressa Aurea" was examined against different leukemia cell lines.

Materials And Methods: Ultra-performance liquid chromatography (UPLC) analysis was performed with the use of a Waters ACQUITY UPLC(TM) system (Waters Corp., Milford, MA). The peaks of podophyllotoxin and deoxypodophyllotoxin were assigned on the basis of their retention data and mass-to-charge ratio (m/z). Trypan blue assay was performed to obtain IC50 cytotoxicity values against selected leukemia cell lines.

Results: Juniperus scopulorum was characterized with the highest level of podophyllotoxin (486.7 mg/100 g DW) while Juniperus davurica Pall. contained the highest amount of deoxypodophyllotoxin (726.8 mg/100 g DW). Podophyllotoxin IC50 cytotoxicity values against J45.01 and CEM/C1 leukemia cell lines were 0.0040 and 0.0286 µg/mL, respectively. Juniperus scopulorum extract examined against J45.01 and HL-60/MX2 leukemia cell lines gave the respective IC50 values: 0.369-9.225 µg/mL. Juniperus communis extract was characterized with the following IC50 cytotoxity values against J45.01 and U-266B1 cell lines: 3.310-24.825 µg/mL.

Conclusions: Juniperus sp. can be considered as an alternative source of podophyllotoxin and deoxypodophyllotoxin. Cytotoxic activity of podophyllotoxin and selected leaf extracts of Juniperus sp. against a set of leukemia cell lines was demonstrated.
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http://dx.doi.org/10.3109/13880209.2014.943246DOI Listing
June 2015

Prevention of tick bites and protection against tick-borne diseases in south-eastern Poland.

Ann Agric Environ Med 2008 ;15(2):181-5

Chair and Department of Biology and Parasitology, Medical University of Lublin, Radziwillowska 11, Lublin, Poland.

The objective of this survey was to obtain information on the level of knowledge of prophylactic behaviour as a protection against fundamental tick-borne diseases among inhabitants of south-eastern Poland, and to determine whether the professions that increase the probability of tick contact, as well as previous contact with ticks, influenced the knowledge on tick-borne diseases and prophylactic behaviour of the respondents. The questionnaire included questions about gender of respondents, their place of residence and preventive behaviour against tick bite. The respondents were asked about the occupational hazard of working in tick infested areas, history of tick bites, awareness of diseases transmitted by ticks and the sources of their knowledge. The research was carried out in 2005. The most frequent preventive measures used were tick repellents (38 % ) and wearing protective clothing (35 % ). According to the questionnaire survey, 21 % of respondents did not take any preventive measures. The practice and form of prophylaxis was associated with the place of residence. In contrast to individuals from rural areas, a relatively low percentage of urban inhabitants used simple methods for protecting themselves against tick bites. Our survey indicates that the awareness of the Lublin region inhabitants concerning methods of tick bite prevention is still low and greater efforts are needed in promoting basic methods of protection and disseminating knowledge about these arthropods.
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March 2009

[The public health risk of ticks and tick-borne diseases in the south-eastern Poland].

Wiad Parazytol 2004 ;50(2):249-52

Katedra i Zakład Biologii i Parazytologii Wydziału Lekarskiego, Akademia Medyczna im. Prof. Feliksa Skubiszewskiego, ul. Radziwillowska 11, 20-080 Lublin.

The aim of the study was to estimate risk of tick-borne diseases among inhabitants of Lublin macroregion. The incidence of Lyme disease in the residents of Lubelskie Voivodeship increased from 2.9 to 6.3 in the period between 2002 and 2003. In 16% of patients studied there were systemic reactions observed after tick bite, while the local symptoms were noted in 56% of bitten subjects. Our data indicates that 59% of the respondents have poor knowledge of ticks and tic-borne diseases.
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November 2006
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