Latest Publications

@article{hyzdalova_transcriptional_2024,

title = {Transcriptional and phenotypical alterations associated with a gradual benzo[a]pyrene-induced transition of human bronchial epithelial cells into mesenchymal-like cells.},

author = {Martina Hýžďalová and Jiřina Procházková and Nicol Straková and Kateřina Pěnčíková and Simona Strapáčová and Jana Slováčková and Simona Kajabová and Helena Líbalová and Jan Topinka and Markéta Kabátková and Jan Vondráček and Steen Mollerup and Miroslav Machala},

doi = {10.1016/j.etap.2024.104424},

issn = {1872-7077 1382-6689},

year  = {2024},

date = {2024-04-01},

journal = {Environmental toxicology and pharmacology},

volume = {107},

pages = {104424},

abstract = {The role of benzo[a]pyrene (BaP), a prominent genotoxic carcinogen and aryl hydrocarbon receptor (AhR) ligand, in tumor progression remains poorly characterized. We investigated the impact of BaP on the process of epithelial-mesenchymal transition (EMT) in normal human bronchial epithelial HBEC-12KT cells. Early morphological changes after 2-week exposure were accompanied with induction of SERPINB2, IL1, CDKN1A/p21 (linked with cell cycle delay) and chemokine CXCL5. After 8-week exposure, induction of cell migration and EMT-related pattern of markers/regulators led to induction of further pro-inflammatory cytokines or non-canonical Wnt pathway ligand WNT5A. This trend of up-regulation of pro-inflammatory genes and non-canonical Wnt pathway constituents was observed also in the BaP-transformed HBEC-12KT-B1 cells. In general, transcriptional effects of BaP differed from those of TGFβ1, a prototypical EMT inducer, or a model non-genotoxic AhR ligand, TCDD. Carcinogenic polycyclic aromatic hydrocarbons could thus induce a unique set of molecular changes linked with EMT and cancer progression.},

note = {Place: Netherlands},

keywords = {*Benzo(a)pyrene/toxicity, *Epithelial Cells/metabolism, 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin, Aryl hydrocarbon receptor, Aryl Hydrocarbon/genetics/metabolism, Benzo[a]pyrene, DNA Damage, Epithelial-Mesenchymal Transition, Human bronchial epithelial cells, Humans, Ligands, Receptors},

pubstate = {published},

tppubtype = {article}

}

@article{vondracek_role_2021,

title = {The Role of Metabolism in Toxicity of Polycyclic Aromatic Hydrocarbons and their Non-genotoxic Modes of Action.},

author = {Jan Vondráček and Miroslav Machala},

doi = {10.2174/1389200221999201125205725},

issn = {1875-5453 1389-2002},

year  = {2021},

date = {2021-01-01},

journal = {Current drug metabolism},

volume = {22},

number = {8},

pages = {584–595},

abstract = {Polycyclic aromatic hydrocarbons (PAHs) represent a class of widely distributed environmental pollutants that have been primarily studied as genotoxic compounds. Their mutagenicity/genotoxicity largely depends on their oxidative metabolism leading to the production of dihydrodiol epoxide metabolites, as well as additional metabolites contributing to oxidative DNA damage, such as PAH quinones. However, both parental PAHs and their metabolites, including PAH quinones or hydroxylated PAHs, have been shown to produce various types of non-genotoxic effects. These include e.g., activation of the aryl hydrocarbon receptor and/or additional nuclear receptors, activation of membrane receptors, including tyrosine kinases and G-protein coupled receptors, or activation of intracellular signaling pathways, such as mitogen-activated protein kinases, Akt kinase and Ca(2+)-dependent signaling. These pathways may, together with the cellular DNA damage responses, modulate cell proliferation, cell survival or cell-to-cell communication, thus contributing to the known carcinogenic effects of PAHs. In the present review, we summarize some of the known non-genotoxic effects of PAHs, focusing primarily on those that have also been shown to be modulated by PAH metabolites. Despite the limitations of the available data, it seems evident that more attention should be paid to the discrimination between the potential non-genotoxic effects of parental PAHs and those of their metabolites. This may provide further insight into the mechanisms of toxicity of this large and diverse group of environmental pollutants.},

note = {Place: Netherlands},

keywords = {Activation, AhR, Animals, Benzo[a]pyrene, Cell Proliferation, Cell Survival, cell-to-cell communication, DNA Damage, Environmental Pollutants/*pharmacokinetics/*toxicity, Humans, Metabolic, Mutagens/*pharmacokinetics/*toxicity, oxidative stress, PAH metabolism., Polycyclic Aromatic Hydrocarbons/*pharmacokinetics/*toxicity},

pubstate = {published},

tppubtype = {article}

}

@article{tylichova_n-3_2019,

title = {n-3 Polyunsaturated fatty acids alter benzo[a]pyrene metabolism and genotoxicity in human colon epithelial cell models.},

author = {Zuzana Tylichová and Jiří Neča and Jan Topinka and Alena Milcová and Jiřina Hofmanová and Alois Kozubík and Miroslav Machala and Jan Vondráček},

doi = {10.1016/j.fct.2018.12.021},

issn = {1873-6351 0278-6915},

year  = {2019},

date = {2019-02-01},

journal = {Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association},

volume = {124},

pages = {374–384},

abstract = {Dietary carcinogens, such as benzo[a]pyrene (BaP), are suspected to contribute to colorectal cancer development. n-3 Polyunsaturated fatty acids (PUFAs) decrease colorectal cancer risk in individuals consuming diets rich in PUFAs. Here, we investigated the impact of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid on metabolism and genotoxicity of BaP in human cell models derived from the colon: HT-29 and HCT-116 cell lines. Both PUFAs reduced levels of excreted BaP metabolites, in particular BaP-tetrols and hydroxylated BaP metabolites, as well as formation of DNA adducts in HT-29 and HCT-116 cells. However, EPA appeared to be a more potent inhibitor of formation of some intracellular BaP metabolites, including BaP-7,8-dihydrodiol. EPA also reduced phosphorylation of histone H2AX (Ser139) in HT-29 cells, which indicated that it may reduce further forms of DNA damage, including DNA double strand breaks. Both PUFAs inhibited induction of CYP1 activity in colon cells determined as 7-ethoxyresorufin-O-deethylase (EROD); this was at least partly linked with inhibition of induction of CYP1A1, 1A2 and 1B1 mRNAs. The downregulation and/or inhibition of CYP1 enzymes by PUFAs could thus alter metabolism and reduce genotoxicity of BaP in human colon cells, which might contribute to known chemopreventive effects of PUFAs in colon epithelium.},

note = {Place: England},

keywords = {Anticarcinogenic Agents/*pharmacology, Benzo(a)pyrene/adverse effects/*metabolism, Cell Line, Colon cancer, Cytochrome P450 Family 1/metabolism, DNA Adducts/metabolism, DNA Damage, DNA Damage/drug effects, Docosahexaenoic acid, Docosahexaenoic Acids/*pharmacology, Eicosapentaenoic acid, Eicosapentaenoic Acid/*pharmacology, Epithelial Cells/*drug effects, Histones/metabolism, Humans, Mutagens/adverse effects/*metabolism, Polycyclic aromatic hydrocarbon, S Phase Cell Cycle Checkpoints/drug effects, Tumor},

pubstate = {published},

tppubtype = {article}

}

@article{simeckova_multiparameter_2018,

title = {Multiparameter cytometric analysis of complex cellular response.},

author = {Šárka Šimečková and Radek Fedr and Ján Remšík and Zuzana Kahounová and Eva Slabáková and Karel Souček},

doi = {10.1002/cyto.a.23295},

issn = {1552-4930 1552-4922},

year  = {2018},

date = {2018-02-01},

journal = {Cytometry. Part A : the journal of the International Society for Analytical Cytology},

volume = {93},

number = {2},

pages = {239–248},

abstract = {Complex analysis of cellular responses after experimental treatment is important for screening, mechanistic understanding of treatment effects, and the identification of sensitive and resistant cell phenotypes. Modern multicolor flow cytometry has demonstrated its power for such analyses. Here, we introduce a multiparametric protocol for complex analysis of cytokinetics by the simultaneous detection of seven fluorescence parameters. This analysis includes the detection of two surface markers for immunophenotyping, analysis of proliferation based on the cell cycle and the measurement of incorporated nucleoside analogue 5-ethynyl-2'-deoxyuridine (EdU) in newly synthesized DNA, analysis of DNA damage using an anti-phospho-histone H2A.X (Ser139) antibody, and determination of cell death using a fixable viability probe and intracellular detection of caspase-3 activation. To demonstrate the applicability of this protocol for the analysis of heterogeneous and complex cell responses, we used different treatments and model cell lines. We demonstrated that this protocol has the potential to provide complex and simultaneous analysis of cytokinetics and analyze the heterogeneity of the response at the single-cell level. © 2017 International Society for Advancement of Cytometry.},

note = {Place: United States},

keywords = {Apoptosis, Apoptosis/*physiology, Cell Line, Cell Proliferation/*physiology, DNA Damage, DNA Damage/*physiology, Flow Cytometry, Flow Cytometry/*methods, Humans, immunophenotyping, Immunophenotyping/*methods, multiparametric analysis, proliferation, Tumor},

pubstate = {published},

tppubtype = {article}

}

@article{palkova_aryl_2015,

title = {The aryl hydrocarbon receptor-mediated and genotoxic effects of fractionated extract of standard reference diesel exhaust particle material in pulmonary, liver and prostate cells.},

author = {Lenka Pálková and Jan Vondráček and Lenka Trilecová and Miroslav Ciganek and Kateřina Pěnčíková and Jiří Neča and Alena Milcová and Jan Topinka and Miroslav Machala},

doi = {10.1016/j.tiv.2014.12.002},

issn = {1879-3177 0887-2333},

year  = {2015},

date = {2015-04-01},

journal = {Toxicology in vitro : an international journal published in association with BIBRA},

volume = {29},

number = {3},

pages = {438–448},

abstract = {Diesel exhaust particles (DEP) and the associated complex mixtures of organic pollutants, such as polycyclic aromatic hydrocarbons (PAHs), or their derivatives, have been suggested to exert deleterious effects on human health. We used a set of defined cellular models representing liver, lung and prostate tissues, in order to compare non-genotoxic and genotoxic effects of crude and fractionated extract of a standard reference DEP material - SRM 1650b. We focused on the aryl hydrocarbon receptor (AhR)-mediated activity, modulation of cell proliferation, formation of DNA adducts, oxidative DNA damage, and induction of DNA damage responses, including evaluation of apoptosis, and phosphorylation of p53 tumor suppressor and checkpoint kinases (Chk). Both PAHs and the polar aromatic compounds contributed to the AhR-mediated activity of DEP-associated organic pollutants. The principal identified AhR agonists included benzo[k]fluoranthene, indeno[1,2,3-c,d]pyrene, chrysene and several non-priority PAHs, including benzochrysenes and methylated PAHs. In contrast to PAHs, polar compounds contributed more significantly to overall formation of DNA adducts associated with phosphorylation of p53, Chk1 or Chk2, and partly with apoptosis. Therefore, more attention should be paid to identification of DEP-associated polar organic compounds, contributing to the AhR activation and cytotoxic/genotoxic effects of complex airborne mixtures of organic contaminants produced by diesel engines.},

note = {Place: England},

keywords = {Air Pollutants/*toxicity, Air pollution, Animals, Apoptosis, Apoptosis/drug effects, Aryl Hydrocarbon/*drug effects, Cell Cycle/drug effects, Cell Death/drug effects, Cell Proliferation, DNA adducts, DNA Damage, DNA damage response, Liver/*pathology, Lung/*pathology, Male, Mutagens/*toxicity, PAHs, Particulate Matter/*toxicity, Prostate/*pathology, Rats, Receptors, SRM 1650b, Vehicle Emissions/*toxicity},

pubstate = {published},

tppubtype = {article}

}

@article{benes_inhibition_2011,

title = {Inhibition of topoisomerase IIα: novel function of wedelolactone.},

author = {Petr Benes and Lucia Knopfova and Filip Trcka and Alice Nemajerova and Diana Pinheiro and Karel Soucek and Miroslav Fojta and Jan Smarda},

doi = {10.1016/j.canlet.2011.01.002},

issn = {1872-7980 0304-3835},

year  = {2011},

date = {2011-04-01},

journal = {Cancer letters},

volume = {303},

number = {1},

pages = {29–38},

abstract = {The naturally occurring coumestan wedelolactone has been previously shown to reduce growth of various cancer cells. So far, the growth-suppressing effect of wedelolactone has been attributed to the inhibition of the NFκB transcription factor and/or androgen receptors. We found that wedelolactone suppressed growth and induced apoptosis of androgen receptor-negative MDA-MB-231 breast cancer cells at concentrations that did not inhibit the NFκB activity. The cells responded to wedelolactone by the S and G2/M phase cell cycle arrest and induction of the DNA damage signaling. Wedelolactone interacted with dsDNA and inhibited the activity of DNA topoisomerase IIα. We conclude that wedelolactone can act as growth suppressor independently of NFκB and androgen receptors.},

note = {Place: Ireland},

keywords = {Antigens, Antineoplastic Agents/pharmacology, Apoptosis/drug effects, Breast Neoplasms/*drug therapy/enzymology/pathology, Cell Cycle/drug effects, Cell Growth Processes/drug effects, Cell Line, Cell Survival/drug effects, Coumarins/*pharmacology, DNA Damage, DNA Topoisomerases, DNA-Binding Proteins/*antagonists & inhibitors/metabolism, Enzyme-Linked Immunosorbent Assay, Female, Humans, Immunoblotting, Neoplasm/metabolism, Signal Transduction, Topoisomerase Inhibitors/*pharmacology, Tumor, Type II/metabolism},

pubstate = {published},

tppubtype = {article}

}

@article{soucek_fetal_2010,

title = {Fetal colon cell line FHC exhibits tumorigenic phenotype, complex karyotype, and TP53 gene mutation.},

author = {Karel Soucek and Pavla Gajdusková and Marie Brázdová and Martina Hýzd'alová and Lenka Kocí and David Vydra and Radek Trojanec and Zuzana Pernicová and Lenka Lentvorská and Marián Hajdúch and Jirina Hofmanová and Alois Kozubík},

doi = {10.1016/j.cancergencyto.2009.11.009},

issn = {1873-4456 0165-4608},

year  = {2010},

date = {2010-03-01},

journal = {Cancer genetics and cytogenetics},

volume = {197},

number = {2},

pages = {107–116},

abstract = {Stable cell lines obtained by spontaneous immortalization might represent early stages of malignant transformation and be useful experimental models for studies of mechanisms of cancer development. The FHC (fetal human cells) cell line has been established from normal fetal colonic mucosa. Detailed characterization of this cell line and mechanism of spontaneously acquired immortality have not been described yet. Therefore, we characterized the FHC cell line in terms of its tumorigenicity, cytogenetics, and TP53 gene mutation analysis. FHC cells displayed capability for anchorage-independent growth in semisolid media in vitro and formed solid tumors after transplantation into SCID (severe combined immunodeficiency) mice. This tumorigenic phenotype was associated with hypotriploidy and chromosome number ranging from 66 to 69. Results of comparative genetic hybridization arrays showed that most chromosomes included regions of copy number gains or losses. Region 8q23 approximately 8q24.3 (containing, e.g., MYC proto-oncogene) was present in more than 20 copies per nucleus. Moreover, we identified mutation of TP53 gene in codon 273; triplet CGT coding Arg was changed to CAG coding His. Expression of Pro codon 72 polymorphic variant of p53 was also detected. Mutation of TP53 gene was associated with abolished induction of p21(Waf1/Cip1) and MDM-2 proteins and resistance to apoptosis after genotoxic treatment. Because of their origin from normal fetal colon and their relative resistance to the induction of apoptosis, FHC cells can be considered a valuable experimental model for various studies.},

note = {Place: United States},

keywords = {*Genes, Animals, Apoptosis/physiology, Carcinoembryonic Antigen/metabolism, Cell Adhesion/physiology, Cell Growth Processes/physiology, Cell Line, Cell Transformation, Colon/cytology/metabolism/*physiology, Colonic Neoplasms/*genetics/*pathology, Comparative Genomic Hybridization, Cytogenetic Analysis/methods, DNA Damage, DNA Mutational Analysis/methods, Female, Fetus/cytology, Fluorescence, HCT116 Cells, Humans, In Situ Hybridization, Karyotyping, Keratins/metabolism, Mice, Neoplasm Transplantation, Neoplastic/genetics/pathology, p53, Phenotype, Proto-Oncogene Mas, SCID, Signal Transduction, Transformed},

pubstate = {published},

tppubtype = {article}

}