Latest Publications

@article{rihova_transcription_2022,

title = {Transcription factor c-Myb: novel prognostic factor in osteosarcoma.},

author = {Kamila Říhová and Monika Dúcka and Iva Staniczková Zambo and Ladislava Vymětalová and Martin Šrámek and Filip Trčka and Jan Verner and Stanislav Drápela and Radek Fedr and Tereza Suchánková and Barbora Pavlatovská and Eva Ondroušková and Irena Kubelková and Danica Zapletalová and Štěpán Tuček and Peter Múdry and Dagmar Adámková Krákorová and Lucia Knopfová and Jan Šmarda and Karel Souček and Lubor Borsig and Petr Beneš},

doi = {10.1007/s10585-021-10145-4},

issn = {1573-7276 0262-0898},

year  = {2022},

date = {2022-04-01},

journal = {Clinical & experimental metastasis},

volume = {39},

number = {2},

pages = {375–390},

abstract = {The transcription factor c-Myb is an oncoprotein promoting cell proliferation and survival when aberrantly activated/expressed, thus contributing to malignant transformation. Overexpression of c-Myb has been found in leukemias, breast, colon and adenoid cystic carcinoma. Recent studies revealed its expression also in osteosarcoma cell lines and suggested its functional importance during bone development. However, the relevance of c-Myb in control of osteosarcoma progression remains unknown. A retrospective clinical study was carried out to assess a relationship between c-Myb expression in archival osteosarcoma tissues and prognosis in a cohort of high-grade osteosarcoma patients. In addition, MYB was depleted in metastatic osteosarcoma cell lines SAOS-2 LM5 and 143B and their growth, chemosensitivity, migration and metastatic activity were determined. Immunohistochemical analysis revealed that high c-Myb expression was significantly associated with poor overall survival in the cohort and metastatic progression in young patients. Increased level of c-Myb was detected in metastatic osteosarcoma cell lines and its depletion suppressed their growth, colony-forming capacity, migration and chemoresistance in vitro in a cell line-dependent manner. MYB knock-out resulted in reduced metastatic activity of both SAOS-2 LM5 and 143B cell lines in immunodeficient mice. Transcriptomic analysis revealed the c-Myb-driven functional programs enriched for genes involved in the regulation of cell growth, stress response, cell adhesion and cell differentiation/morphogenesis. Wnt signaling pathway was identified as c-Myb target in osteosarcoma cells. Taken together, we identified c-Myb as a negative prognostic factor in osteosarcoma and showed its involvement in the regulation of osteosarcoma cell growth, chemosensitivity, migration and metastatic activity.},

note = {Place: Netherlands},

keywords = {*Bone Neoplasms/pathology, *Osteosarcoma/pathology, Animals, c-Myb, Cell Line, Cell Movement/genetics, Cell Proliferation, Chemoresistance, Gene Expression Regulation, Humans, Metastasis, Mice, Neoplastic, Osteosarcoma, Prognosis, proliferation, Retrospective Studies, Tumor, Wnt Signaling Pathway},

pubstate = {published},

tppubtype = {article}

}

@article{krkoska_role_2022,

title = {Role of miR-653 and miR-29c in downregulation of CYP1A2 expression in hepatocellular carcinoma.},

author = {Martin Krkoška and Jana Nekvindová and Kateřina Nevědělová and Veronika Zubáňová and Lenka Radová and Jan Vondráček and Jarmila Herůdková and Ondřej Slabý and Igor Kiss and Lucia Bohovicová and Pavel Fabian and Zuzana Tylichová and Zdeněk Kala and Petr Kysela and Lenka Ostřížková and Vladimír Palička and Alena Hyršlová Vaculová},

doi = {10.1007/s43440-021-00338-9},

issn = {2299-5684 1734-1140},

year  = {2022},

date = {2022-02-01},

journal = {Pharmacological reports : PR},

volume = {74},

number = {1},

pages = {148–158},

abstract = {BACKGROUND: Hepatocellular carcinoma (HCC) is a major contributor to the worldwide cancer burden. Recent studies on HCC have demonstrated dramatic alterations in expression of several cytochrome P450 (CYP) family members that play a crucial role in biotransformation of many drugs and other xenobiotics; however, the mechanisms responsible for their deregulation remain unclear. METHODS: We investigated a potential involvement of miRNAs in downregulation of expression of CYPs observed in HCC tumors. We compared miRNA expression profiles (TaqMan Array Human MicroRNA v3.0 TLDA qPCR) between HCC human patient tumors with strong (CYP-) and weak/no (CYP+) downregulation of drug-metabolizing CYPs. The role of significantly deregulated miRNAs in modulation of expression of the CYPs and associated xenobiotic receptors was then investigated in human liver HepaRG cells transfected with relevant miRNA mimics or inhibitors. RESULTS: We identified five differentially expressed miRNAs in CYP- versus CYP+ tumors, namely miR-29c, miR-125b1, miR-505, miR-653 and miR-675. The two most-upregulated miRNAs found in CYP- tumor samples, miR-29c and miR-653, were found to act as efficient suppressors of CYP1A2 or AHR expression. CONCLUSIONS: Our results revealed a novel role of miR-653 and miR-29c in regulation of expresion of CYPs involved in crucial biotransformation processes in liver, which are often deregulated during liver cancer progression.},

note = {Place: Switzerland},

keywords = {*Carcinoma, *Liver Neoplasms/genetics/metabolism, AhR, Biotransformation, Cell Line, CYP1A2, Cytochrome P-450 CYP1A2/*metabolism, Down-Regulation, Gene Expression Regulation, Hepatocellular carcinoma, Hepatocellular/genetics/metabolism, Hepatocytes/metabolism, Humans, MicroRNA, MicroRNAs/*metabolism, Neoplastic, Tumor, Xenobiotics/metabolism},

pubstate = {published},

tppubtype = {article}

}

@article{krkoska_deregulation_2021,

title = {Deregulation of signaling pathways controlling cell survival and proliferation in cancer cells alters induction of cytochrome P450 family 1 enzymes.},

author = {Martin Krkoška and Jana Svobodová and Markéta Kabátková and Ondřej Zapletal and Alena Hyršlová Vaculová and Jana Nekvindová and Jan Vondráček},

doi = {10.1016/j.tox.2021.152897},

issn = {1879-3185 0300-483X},

year  = {2021},

date = {2021-09-01},

journal = {Toxicology},

volume = {461},

pages = {152897},

abstract = {Cytochrome P450 family 1 (CYP1) enzymes contribute both to metabolism of xenobiotics and to the control of endogenous levels of ligands of the aryl hydrocarbon receptor (AhR). Their activities, similar to other CYPs, can be altered in tumor tissues. Here, we examined a possible role of proliferative/survival pathways signaling, which is often deregulated in tumor cells, and possible links with p300 histone acetyltransferase (a transcriptional co-activator) in the control of CYP1 expression, focusing particularly on CYP1A1. Using cell models derived from human liver, we observed that the induction of CYP1A1 expression, as well as other CYP1 enzymes, was reduced in exponentially growing cells, as compared with their non-dividing counterparts. The siRNA-mediated inhibition of proliferation/pro-survival signaling pathway effectors (such as β-catenin and/or Hippo pathway effectors YAP/TAZ) increased the AhR ligand-induced CYP1A1 mRNA levels in liver HepaRG cells, and/or in colon carcinoma HCT-116 cells. The activation of proliferative Wnt/β-catenin signaling in HCT-116 cells reduced both the induction of CYP1 enzymes and the binding of p300 to the promoter of CYP1A1 or CYP1B1 genes. These results seem to indicate that aberrant proliferative signaling in tumor cells could suppress induction of CYP1A1 (or other CYP1 enzymes) via competition for p300 binding. This mechanism could be involved in modulation of the metabolism of both endogenous and exogenous substrates of CYP1A1 (and other CYP1 enzymes), with possible further consequences for alterations of the AhR signaling in tumor cells, or additional functional roles of CYP1 enzymes.},

note = {Place: Ireland},

keywords = {AhR, Cancer cells, Cell Line, Cell Proliferation, Cell Proliferation/*physiology, Cell Survival/physiology, Colonic Neoplasms/genetics/*pathology, CYP1 enzymes, Cytochrome P-450 CYP1A1/biosynthesis/*genetics, E1A-Associated p300 Protein/metabolism, Enzyme Induction/physiology, Gene Expression Regulation, HCT116 Cells, Hippo Signaling Pathway/physiology, Humans, Liver/*pathology, Neoplastic, p300, Signal Transduction/physiology, Tumor, Wnt Signaling Pathway/physiology, β-Catenin signaling},

pubstate = {published},

tppubtype = {article}

}

@article{machala_changes_2021,

title = {Changes in Sphingolipid Profile of Benzo[a]pyrene-Transformed Human Bronchial Epithelial Cells Are Reflected in the Altered Composition of Sphingolipids in Their Exosomes.},

author = {Miroslav Machala and Josef Slavík and Ondrej Kováč and Jiřina Procházková and Kateřina Pěnčíková and Martina Pařenicová and Nicol Straková and Jan Kotouček and Pavel Kulich and Steen Mollerup and Jan Vondráček and Martina Hýžďalová},

doi = {10.3390/ijms22179195},

issn = {1422-0067},

year  = {2021},

date = {2021-08-01},

journal = {International journal of molecular sciences},

volume = {22},

number = {17},

abstract = {Sphingolipids (SLs), glycosphingolipids (GSLs), and eicosanoids are bioactive lipids, which play important roles in the etiology of various diseases, including cancer. However, their content and roles in cancer cells, and in particular in the exosomes derived from tumor cells, remain insufficiently characterized. In this study, we evaluated alterations of SL and GSL levels in transformed cells and their exosomes, using comparative HPLC-MS/MS analysis of parental human bronchial epithelial cells HBEC-12KT and their derivative, benzo[a]pyrene-transformed HBEC-12KT-B1 cells with the acquired mesenchymal phenotype. We examined in parallel SL/GSL contents in the exosomes released from both cell lines. We found significant alterations of the SL/GSL profile in the transformed cell line, which corresponded well with alterations of the SL/GSL profile in exosomes derived from these cells. This suggested that a majority of SLs and GSLs were transported by exosomes in the same relative pattern as in the cells of origin. The only exceptions included decreased contents of sphingosin, sphingosin-1-phosphate, and lactosylceramide in exosomes derived from the transformed cells, as compared with the exosomes derived from the parental cell line. Importantly, we found increased levels of ceramide phosphate, globoside Gb3, and ganglioside GD3 in the exosomes derived from the transformed cells. These positive modulators of epithelial-mesenchymal transition and other pro-carcinogenic processes might thus also contribute to cancer progression in recipient cells. In addition, the transformed HBEC-12KT-B1 cells also produced increased amounts of eicosanoids, in particular prostaglandin E2. Taken together, the exosomes derived from the transformed cells with specifically upregulated SL and GSL species, and increased levels of eicosanoids, might contribute to changes within the cancer microenvironment and in recipient cells, which could in turn participate in cancer development. Future studies should address specific roles of individual SL and GSL species identified in the present study.},

note = {Place: Switzerland},

keywords = {*Cell Transformation, Benzo(a)pyrene/toxicity, Bronchi/cytology, Carcinogens/toxicity, Cell Line, eicosanoids, exosomes, Exosomes/*metabolism, glycosphingolipid, Humans, in vitro cell transformation, Neoplastic, Respiratory Mucosa/drug effects/*metabolism, sphingolipid, Sphingolipids/*metabolism},

pubstate = {published},

tppubtype = {article}

}

@article{hofmanova_complex_2021,

title = {Complex Alterations of Fatty Acid Metabolism and Phospholipidome Uncovered in Isolated Colon Cancer Epithelial Cells.},

author = {Jiřina Hofmanová and Josef Slavík and Miroslav Ciganek and Petra Ovesná and Zuzana Tylichová and Martina Karasová and Ondřej Zapletal and Nicol Straková and Jiřina Procházková and Jan Bouchal and Zdeněk Kolář and Jiří Ehrmann and Monika Levková and Zlatka Hušková and Pavel Skalický and Alois Kozubík and Miroslav Machala and Jan Vondráček},

doi = {10.3390/ijms22136650},

issn = {1422-0067},

year  = {2021},

date = {2021-06-01},

journal = {International journal of molecular sciences},

volume = {22},

number = {13},

abstract = {The development of colon cancer, one of the most common malignancies, is accompanied with numerous lipid alterations. However, analyses of whole tumor samples may not always provide an accurate description of specific changes occurring directly in tumor epithelial cells. Here, we analyzed in detail the phospholipid (PL), lysophospholipid (lysoPL), and fatty acid (FA) profiles of purified EpCAM(+) cells, isolated from tumor and adjacent non-tumor tissues of colon cancer patients. We found that a number of FAs increased significantly in isolated tumor cells, which also included a number of long polyunsaturated FAs. Higher levels of FAs were associated with increased expression of FA synthesis genes, as well as with altered expression of enzymes involved in FA elongation and desaturation, including particularly fatty acid synthase, stearoyl-CoA desaturase, fatty acid desaturase 2 and ELOVL5 fatty acid elongase 5 We identified significant changes in ratios of specific lysoPLs and corresponding PLs. A number of lysophosphatidylcholine and lysophosphatidylethanolamine species, containing long-chain and very-long chain FAs, often with high numbers of double bonds, were significantly upregulated in tumor cells. Increased de novo synthesis of very long-chain FAs, or, altered uptake or incorporation of these FAs into specific lysoPLs in tumor cells, may thus contribute to reprogramming of cellular phospholipidome and membrane alterations observed in colon cancer.},

note = {Place: Switzerland},

keywords = {*Gene Expression Regulation, *Lipid Metabolism, Adenocarcinoma/enzymology/genetics/*metabolism, Aged, Colonic Neoplasms/enzymology/genetics/*metabolism, colorectal carcinoma, desaturation, EpCAM, Epithelial Cells, Epithelial Cells/enzymology/metabolism, Fatty Acid Desaturases/genetics/metabolism, Fatty Acid Elongases/genetics/metabolism, Fatty Acid Synthases/genetics/metabolism, fatty acid synthesis, Fatty Acids/*metabolism, Female, Humans, lipidomics, Lipogenesis, lysophospholipids, Male, Neoplastic, Phospholipids, Phospholipids/*metabolism, Stearoyl-CoA Desaturase/genetics/metabolism},

pubstate = {published},

tppubtype = {article}

}

@article{mickova_skp2_2021,

title = {Skp2 and Slug Are Coexpressed in Aggressive Prostate Cancer and Inhibited by Neddylation Blockade.},

author = {Alena Mickova and Gvantsa Kharaishvili and Daniela Kurfurstova and Mariam Gachechiladze and Milan Kral and Ondrej Vacek and Barbora Pokryvkova and Martin Mistrik and Karel Soucek and Jan Bouchal},

doi = {10.3390/ijms22062844},

issn = {1422-0067},

year  = {2021},

date = {2021-03-01},

journal = {International journal of molecular sciences},

volume = {22},

number = {6},

abstract = {Prostate cancer (PCa) is the second leading cause of cancer-related deaths in men in Western countries, and there is still an urgent need for a better understanding of PCa progression to inspire new treatment strategies. Skp2 is a substrate-recruiting component of the E3 ubiquitin ligase complex, whose activity is regulated through neddylation. Slug is a transcriptional repressor involved in the epithelial-to-mesenchymal transition, which may contribute to therapy resistance. Although Skp2 has previously been associated with a mesenchymal phenotype and prostate cancer progression, the relationship with Slug deserves further elucidation. We have previously shown that a high Gleason score (≥8) is associated with higher Skp2 and lower E-cadherin expression. In this study, significantly increased expression of Skp2, AR, and Slug, along with E-cadherin downregulation, was observed in primary prostate cancer in patients who already had lymph node metastases. Skp2 was slightly correlated with Slug and AR in the whole cohort (Rs 0.32 and 0.37, respectively), which was enhanced for both proteins in patients with high Gleason scores (Rs 0.56 and 0.53, respectively) and, in the case of Slug, also in patients with metastasis to lymph nodes (Rs 0.56). Coexpression of Skp2 and Slug was confirmed in prostate cancer tissues by multiplex immunohistochemistry and confocal microscopy. The same relationship between these two proteins was observed in three sets of prostate epithelial cell lines (PC3, DU145, and E2) and their mesenchymal counterparts. Chemical inhibition of Skp2, but not RNA interference, modestly decreased Slug protein in PC3 and its docetaxel-resistant subline PC3 DR12. Importantly, chemical inhibition of Skp2 by MLN4924 upregulated p27 and decreased Slug expression in PC3, PC3 DR12, and LAPC4 cells. Novel treatment strategies targeting Skp2 and Slug by the neddylation blockade may be promising in advanced prostate cancer, as recently documented for other aggressive solid tumors.},

note = {Place: Switzerland},

keywords = {*Protein Processing, Androgen/genetics/metabolism, Antigens, Antineoplastic Agents/pharmacology, Cadherins/genetics/metabolism, CD/genetics/metabolism, Cell Line, Cell Survival/drug effects, Cyclin-Dependent Kinase Inhibitor p27/genetics/metabolism, Cyclopentanes/pharmacology, Docetaxel/pharmacology, Epithelial-Mesenchymal Transition/genetics, Gene Expression Regulation, Humans, Immunohistochemistry, Lymphatic Metastasis, Male, multiplex, NEDD8 Protein/*genetics/metabolism, neddylation, Neoplasm Grading, Neoplastic, PC-3 Cells, Post-Translational, Prostate cancer, Prostate/metabolism/pathology, Prostatic Neoplasms/*genetics/metabolism/pathology, Pyrimidines/pharmacology, Receptors, RNA, S-Phase Kinase-Associated Proteins/antagonists & inhibitors/*genetics/metabolism, Skp2 (S-phase kinase-associated protein 2), Slug, Small Interfering/genetics/metabolism, Snail Family Transcription Factors/*genetics/metabolism, Tumor},

pubstate = {published},

tppubtype = {article}

}

@article{remsik_tgf-_2020,

title = {TGF-β regulates Sca-1 expression and plasticity of pre-neoplastic mammary epithelial stem cells.},

author = {Ján Remšík and Markéta Pícková and Ondřej Vacek and Radek Fedr and Lucia Binó and Aleš Hampl and Karel Souček},

doi = {10.1038/s41598-020-67827-4},

issn = {2045-2322},

year  = {2020},

date = {2020-07-01},

journal = {Scientific reports},

volume = {10},

number = {1},

pages = {11396},

abstract = {The epithelial-mesenchymal plasticity, in tight association with stemness, contributes to the mammary gland homeostasis, evolution of early neoplastic lesions and cancer dissemination. Focused on cell surfaceome, we used mouse models of pre-neoplastic mammary epithelial and cancer stem cells to reveal the connection between cell surface markers and distinct cell phenotypes. We mechanistically dissected the TGF-β family-driven regulation of Sca-1, one of the most commonly used adult stem cell markers. We further provided evidence that TGF-β disrupts the lineage commitment and promotes the accumulation of tumor-initiating cells in pre-neoplastic cells.},

note = {Place: England},

keywords = {Animal/pathology, Animals, Ataxin-1/*metabolism, Breast Neoplasms/genetics/*pathology, Cell Line, Cell Plasticity/genetics, Epithelial Cells/pathology, Epithelial-Mesenchymal Transition/genetics, ErbB-2/genetics, Experimental/genetics/*pathology, Female, Gene Expression Regulation, Humans, Mammary Glands, Mammary Neoplasms, Mice, Neoplastic, Neoplastic Stem Cells/*pathology, Receptor, Recombinant Proteins/genetics/metabolism, Signal Transduction/genetics, Transforming Growth Factor beta/genetics/*metabolism, Tumor/transplantation},

pubstate = {published},

tppubtype = {article}

}

@article{machala_colon_2019,

title = {Colon Cancer and Perturbations of the Sphingolipid Metabolism.},

author = {Miroslav Machala and Jiřina Procházková and Jiřina Hofmanová and Lucie Králiková and Josef Slavík and Zuzana Tylichová and Petra Ovesná and Alois Kozubík and Jan Vondráček},

doi = {10.3390/ijms20236051},

issn = {1422-0067},

year  = {2019},

date = {2019-11-01},

journal = {International journal of molecular sciences},

volume = {20},

number = {23},

abstract = {The development and progression of colorectal cancer (CRC), a major cause of cancer-related death in the western world, is accompanied with alterations of sphingolipid (SL) composition in colon tumors. A number of enzymes involved in the SL metabolism have been found to be deregulated in human colon tumors, in experimental rodent studies, and in human colon cancer cells in vitro. Therefore, the enzymatic pathways that modulate SL levels have received a significant attention, due to their possible contribution to CRC development, or as potential therapeutic targets. Many of these enzymes are associated with an increased sphingosine-1-phosphate/ceramide ratio, which is in turn linked with increased colon cancer cell survival, proliferation and cancer progression. Nevertheless, more attention should also be paid to the more complex SLs, including specific glycosphingolipids, such as lactosylceramides, which can be also deregulated during CRC development. In this review, we focus on the potential roles of individual SLs/SL metabolism enzymes in colon cancer, as well as on the pros and cons of employing the current in vitro models of colon cancer cells for lipidomic studies investigating the SL metabolism in CRC.},

note = {Place: Switzerland},

keywords = {*Gene Expression Regulation, Acid Ceramidase/genetics/metabolism, Alkaline Ceramidase/genetics/metabolism, Animal, Animals, Ceramides/metabolism, colon cancer (CRC) sphingolipidomics, colon cancer cells, Colonic Neoplasms/*enzymology/genetics/pathology, colorectal cancer, Cultured, Disease Models, glycosphingolipid, Humans, Lactosylceramide, Lactosylceramides/*metabolism, Lipid Metabolism/*genetics, Lysophospholipids/metabolism, Neoplastic, Neutral Ceramidase/genetics/metabolism, Phosphotransferases (Alcohol Group Acceptor)/genetics/metabolism, Proto-Oncogene Proteins c-akt/genetics/metabolism, sphingolipid, Sphingolipids/*metabolism, Sphingosine N-Acyltransferase/genetics/metabolism, sphingosine-1-phosphate, Sphingosine/analogs & derivatives/metabolism, Tumor Cells},

pubstate = {published},

tppubtype = {article}

}

@article{simeckova_high_2019,

title = {High Skp2 expression is associated with a mesenchymal phenotype and increased tumorigenic potential of prostate cancer cells.},

author = {Šárka Šimečková and Zuzana Kahounová and Radek Fedr and Ján Remšík and Eva Slabáková and Tereza Suchánková and Jiřina Procházková and Jan Bouchal and Gvantsa Kharaishvili and Milan Král and Petr Beneš and Karel Souček},

doi = {10.1038/s41598-019-42131-y},

issn = {2045-2322},

year  = {2019},

date = {2019-04-01},

journal = {Scientific reports},

volume = {9},

number = {1},

pages = {5695},

abstract = {Skp2 is a crucial component of SCF(Skp2) E3 ubiquitin ligase and is often overexpressed in various types of cancer, including prostate cancer (PCa). The epithelial-to-mesenchymal transition (EMT) is involved in PCa progression. The acquisition of a mesenchymal phenotype that results in a cancer stem cell (CSC) phenotype in PCa was described. Therefore, we aimed to investigate the expression and localization of Skp2 in clinical samples from patients with PCa, the association of Skp2 with EMT status, and the role of Skp2 in prostate CSC. We found that nuclear expression of Skp2 was increased in patients with PCa compared to those with benign hyperplasia, and correlated with high Gleason score in PCa patients. Increased Skp2 expression was observed in PCa cell lines with mesenchymal and CSC-like phenotype compared to their epithelial counterparts. Conversely, the CSC-like phenotype was diminished in cells in which SKP2 expression was silenced. Furthermore, we observed that Skp2 downregulation led to the decrease in subpopulation of CD44(+)CD24(-) cancer stem-like cells. Finally, we showed that high expression levels of both CD24 and CD44 were associated with favorable recurrence-free survival for PCa patients. This study uncovered the Skp2-mediated CSC-like phenotype with oncogenic functions in PCa.},

note = {Place: England},

keywords = {*Epithelial-Mesenchymal Transition, *Gene Expression Regulation, Animals, CD24 Antigen/genetics, Cell Line, Humans, Hyaluronan Receptors/genetics, Male, Mice, Neoplasm Grading, Neoplastic, Neoplastic Stem Cells/metabolism/*physiology, Nude, PC-3 Cells, Prostatic Neoplasms/*genetics/metabolism/physiopathology, S-Phase Kinase-Associated Proteins/*genetics, Tumor, Xenograft Model Antitumor Assays},

pubstate = {published},

tppubtype = {article}

}