Latest Publications

@article{valcikova_eif4f_2024,

title = {eIF4F controls ERK MAPK signaling in melanomas with BRAF and NRAS mutations.},

author = {Barbora Valcikova and Natalia Vadovicova and Karolina Smolkova and Magdalena Zacpalova and Pavel Krejci and Shannon Lee and Jens Rauch and Walter Kolch and Alexander Kriegsheim and Anna Dorotikova and Zdenek Andrysik and Rachel Vichova and Ondrej Vacek and Karel Soucek and Stjepan Uldrijan},

doi = {10.1073/pnas.2321305121},

issn = {1091-6490 0027-8424},

year  = {2024},

date = {2024-10-01},

journal = {Proceedings of the National Academy of Sciences of the United States of America},

volume = {121},

number = {44},

pages = {e2321305121},

abstract = {The eIF4F translation initiation complex plays a critical role in melanoma resistance to clinical BRAF and MEK inhibitors. In this study, we uncover a function of eIF4F in the negative regulation of the rat sarcoma (RAS)/rapidly accelerated fibrosarcoma (RAF)/mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase (MAPK) signaling pathway. We demonstrate that eIF4F is essential for controlling ERK signaling intensity in treatment-naïve melanoma cells harboring BRAF or NRAS mutations. Specifically, the dual-specificity phosphatase DUSP6/MKP3, which acts as a negative feedback regulator of ERK activity, requires continuous production in an eIF4F-dependent manner to limit excessive ERK signaling driven by oncogenic RAF/RAS mutations. Treatment with small-molecule eIF4F inhibitors disrupts the negative feedback control of MAPK signaling, leading to ERK hyperactivation and EGR1 overexpression in melanoma cells in vitro and in vivo. Furthermore, our quantitative analyses reveal a high spare signaling capacity in the ERK pathway, suggesting that eIF4F-dependent feedback keeps the majority of ERK molecules inactive under normal conditions. Overall, our findings highlight the crucial role of eIF4F in regulating ERK signaling flux and suggest that pharmacological eIF4F inhibitors can disrupt the negative feedback control of MAPK activity in melanomas with BRAF and NRAS activating mutations.},

note = {Place: United States},

keywords = {*Eukaryotic Initiation Factor-4F/metabolism/genetics, *GTP Phosphohydrolases/metabolism/genetics, *MAP Kinase Signaling System/genetics, *Melanoma/genetics/metabolism/pathology, *Membrane Proteins/metabolism/genetics, *Mutation, *Proto-Oncogene Proteins B-raf/genetics/metabolism, Animals, Cell Line, Dual Specificity Phosphatase 6/metabolism/genetics, DUSP6, eIF4F, ERK, Extracellular Signal-Regulated MAP Kinases/metabolism, Humans, MAP kinase, Melanoma, Mice, Tumor},

pubstate = {published},

tppubtype = {article}

}

@article{nemec_discovery_2024,

title = {Discovery of Two Highly Selective Structurally Orthogonal Chemical Probes for Activin Receptor-like Kinases 1 and 2.},

author = {Václav Němec and Marek Remeš and Petr Beňovský and Michael C. Böck and Eliška Šranková and Jong Fu Wong and Julien Cros and Eleanor Williams and Lap Hang Tse and David Smil and Deeba Ensan and Methvin B. Isaac and Rima Al-Awar and Regina Gomolková and Vlad-Constantin Ursachi and Bohumil Fafílek and Zuzana Kahounová and Ráchel Víchová and Ondřej Vacek and Benedict-Tilman Berger and Carrow I. Wells and Cesear R. Corona and James D. Vasta and Matthew B. Robers and Pavel Krejci and Karel Souček and Alex N. Bullock and Stefan Knapp and Kamil Paruch},

doi = {10.1021/acs.jmedchem.4c00629},

issn = {1520-4804 0022-2623},

year  = {2024},

date = {2024-08-01},

journal = {Journal of medicinal chemistry},

volume = {67},

number = {15},

pages = {12632–12659},

abstract = {Activin receptor-like kinases 1-7 (ALK1-7) regulate a complex network of SMAD-independent as well as SMAD-dependent signaling pathways. One of the widely used inhibitors for functional investigations of these processes, in particular for bone morphogenetic protein (BMP) signaling, is LDN-193189. However, LDN-193189 has insufficient kinome-wide selectivity complicating its use in cellular target validation assays. Herein, we report the identification and comprehensive characterization of two chemically distinct highly selective inhibitors of ALK1 and ALK2, M4K2234 and MU1700, along with their negative controls. We show that both MU1700 and M4K2234 efficiently block the BMP pathway via selective in cellulo inhibition of ALK1/2 kinases and exhibit favorable in vivo profiles in mice. MU1700 is highly brain penetrant and shows remarkably high accumulation in the brain. These high-quality orthogonal chemical probes offer the selectivity required to become widely used tools for in vitro and in vivo investigation of BMP signaling.},

note = {Place: United States},

keywords = {*Activin Receptors, Activin Receptors, Animals, Bone Morphogenetic Proteins/metabolism, Drug Discovery, Humans, Mice, Molecular Probes/chemistry, Protein Kinase Inhibitors/pharmacology/chemistry, Pyrazoles/chemistry/pharmacology/chemical synthesis, Signal Transduction/drug effects, Structure-Activity Relationship, Type I/antagonists & inhibitors/metabolism, Type II/metabolism/antagonists & inhibitors},

pubstate = {published},

tppubtype = {article}

}

@article{vazquez-gomez_aryl_2023,

title = {Aryl hydrocarbon receptor (AhR) and pregnane X receptor (PXR) play both distinct and common roles in the regulation of colon homeostasis and intestinal carcinogenesis.},

author = {Gerardo Vázquez-Gómez and Jiří Petráš and Zdeněk Dvořák and Jan Vondráček},

doi = {10.1016/j.bcp.2023.115797},

issn = {1873-2968 0006-2952},

year  = {2023},

date = {2023-10-01},

journal = {Biochemical pharmacology},

volume = {216},

pages = {115797},

abstract = {Both aryl hydrocarbon receptor (AhR) and pregnane X receptor (PXR) belong among key regulators of xenobiotic metabolism in the intestinal tissue. AhR in particular is activated by a wide range of environmental and dietary carcinogens. The data accumulated over the last two decades suggest that both of these transcriptional regulators play a much wider role in the maintenance of gut homeostasis, and that both transcription factors may affect processes linked with intestinal tumorigenesis. Intestinal epithelium is continuously exposed to a wide range of AhR, PXR and dual AhR/PXR ligands formed by intestinal microbiota or originating from diet. Current evidence suggests that specific ligands of both AhR and PXR can protect intestinal epithelium against inflammation and assist in the maintenance of epithelial barrier integrity. AhR, and to a lesser extent also PXR, have been shown to play a protective role against inflammation-induced colon cancer, or, in mouse models employing overactivation of Wnt/β-catenin signaling. In contrast, other evidence suggests that both receptors may contribute to modulation of transformed colon cell behavior, with a potential to promote cancer progression and/or chemoresistance. The review focuses on both overlapping and separate roles of the two receptors in these processes, and on possible implications of their activity within the context of intestinal tissue.},

note = {Place: England},

keywords = {*Receptors, Animals, Aryl hydrocarbon receptor, Aryl Hydrocarbon/genetics/metabolism, Carcinogenesis/genetics/metabolism, Colon cancer, Colon/metabolism, Dietary contaminants, Epithelial barrier, Inflammation, Inflammation/metabolism, Intestine, Mice, Microbial agonists, Pregnane X receptor, Pregnane X Receptor/metabolism, Steroid/metabolism},

pubstate = {published},

tppubtype = {article}

}

@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{radaszkiewicz_rnf43_2021,

title = {RNF43 inhibits WNT5A-driven signaling and suppresses melanoma invasion and resistance to the targeted therapy.},

author = {Tomasz Radaszkiewicz and Michaela Nosková and Kristína Gömöryová and Olga Vondálová Blanářová and Katarzyna Anna Radaszkiewicz and Markéta Picková and Ráchel Víchová and Tomáš Gybeľ and Karol Kaiser and Lucia Demková and Lucia Kučerová and Tomáš Bárta and David Potěšil and Zbyněk Zdráhal and Karel Souček and Vítězslav Bryja},

doi = {10.7554/eLife.65759},

issn = {2050-084X},

year  = {2021},

date = {2021-10-01},

journal = {eLife},

volume = {10},

abstract = {RNF43 is an E3 ubiquitin ligase and known negative regulator of WNT/β-catenin signaling. We demonstrate that RNF43 is also a regulator of noncanonical WNT5A-induced signaling in human cells. Analysis of the RNF43 interactome using BioID and immunoprecipitation showed that RNF43 can interact with the core receptor complex components dedicated to the noncanonical Wnt pathway such as ROR1, ROR2, VANGL1, and VANGL2. RNF43 triggers VANGL2 ubiquitination and proteasomal degradation and clathrin-dependent internalization of ROR1 receptor and inhibits ROR2 activation. These activities of RNF43 are physiologically relevant and block pro-metastatic WNT5A signaling in melanoma. RNF43 inhibits responses to WNT5A, which results in the suppression of invasive properties of melanoma cells. Furthermore, RNF43 prevented WNT5A-assisted development of resistance to BRAF V600E and MEK inhibitors. Next, RNF43 acted as melanoma suppressor and improved response to targeted therapies in vivo. In line with these findings, RNF43 expression decreases during melanoma progression and RNF43-low patients have a worse prognosis. We conclude that RNF43 is a newly discovered negative regulator of WNT5A-mediated biological responses that desensitizes cells to WNT5A.},

note = {Place: England},

keywords = {*Melanoma/genetics/pathology/prevention & control, *Signal Transduction, Animals, BRAF V600E, cancer biology, cell biology, human, Inbred NOD, Male, Melanoma, Mice, mouse, Neoplasm Invasiveness/genetics, RNF43, ROR1, Ubiquitin-Protein Ligases/*genetics/metabolism, VANGL1, Wnt-5a Protein/*genetics/metabolism, WNT5A},

pubstate = {published},

tppubtype = {article}

}

@article{nemec_highly_2021,

title = {Highly selective inhibitors of protein kinases CLK and HIPK with the furo[3,2-b]pyridine core.},

author = {Václav Němec and Lukáš Maier and Benedict-Tilman Berger and Apirat Chaikuad and Stanislav Drápela and Karel Souček and Stefan Knapp and Kamil Paruch},

doi = {10.1016/j.ejmech.2021.113299},

issn = {1768-3254 0223-5234},

year  = {2021},

date = {2021-04-01},

journal = {European journal of medicinal chemistry},

volume = {215},

pages = {113299},

abstract = {The furo [3,2-b]pyridine motif represents a relatively underexplored central pharmacophore in the area of kinase inhibitors. Herein, we report flexible synthesis of 3,5-disubstituted furo [3,2-b]pyridines that relies on chemoselective couplings of newly prepared 5-chloro-3-iodofuro [3,2-b]pyridine. This methodology allowed efficient second-generation synthesis of the state-of-the-art chemical biology probe for CLK1/2/4 MU1210, and identification of the highly selective inhibitors of HIPKs MU135 and MU1787 which are presented and characterized in this study, including the X-ray crystal structure of MU135 in HIPK2. chemical biology probe.},

note = {Place: France},

keywords = {2-b]pyridine, Animals, Carrier Proteins/*antagonists & inhibitors/metabolism, CLK, Crystallography, Furans/chemical synthesis/metabolism/*pharmacology, Furo[3, HIPK, Humans, Inhibitor, Kinase, MCF-7 Cells, Mice, Molecular Structure, MU1210, MU135, MU1787, Protein Binding, Protein Kinase Inhibitors/chemical synthesis/metabolism/*pharmacology, Protein Serine-Threonine Kinases/*antagonists & inhibitors/metabolism, Pyridines/chemical synthesis/metabolism/*pharmacology, Structure-Activity Relationship, X-Ray},

pubstate = {published},

tppubtype = {article}

}

@article{drapela_chk1_2020,

title = {The CHK1 inhibitor MU380 significantly increases the sensitivity of human docetaxel-resistant prostate cancer cells to gemcitabine through the induction of mitotic catastrophe.},

author = {Stanislav Drápela and Prashant Khirsariya and Wytske M. Weerden and Radek Fedr and Tereza Suchánková and Diana Búzová and Jan Červený and Aleš Hampl and Martin Puhr and William R. Watson and Zoran Culig and Lumír Krejčí and Kamil Paruch and Karel Souček},

doi = {10.1002/1878-0261.12756},

issn = {1878-0261 1574-7891},

year  = {2020},

date = {2020-10-01},

journal = {Molecular oncology},

volume = {14},

number = {10},

pages = {2487–2503},

abstract = {As treatment options for patients with incurable metastatic castration-resistant prostate cancer (mCRPC) are considerably limited, novel effective therapeutic options are needed. Checkpoint kinase 1 (CHK1) is a highly conserved protein kinase implicated in the DNA damage response (DDR) pathway that prevents the accumulation of DNA damage and controls regular genome duplication. CHK1 has been associated with prostate cancer (PCa) induction, progression, and lethality; hence, CHK1 inhibitors SCH900776 (also known as MK-8776) and the more effective SCH900776 analog MU380 may have clinical applications in the therapy of PCa. Synergistic induction of DNA damage with CHK1 inhibition represents a promising therapeutic approach that has been tested in many types of malignancies, but not in chemoresistant mCRPC. Here, we report that such therapeutic approach may be exploited using the synergistic action of the antimetabolite gemcitabine (GEM) and CHK1 inhibitors SCH900776 and MU380 in docetaxel-resistant (DR) mCRPC. Given the results, both CHK1 inhibitors significantly potentiated the sensitivity to GEM in a panel of chemo-naïve and matched DR PCa cell lines under 2D conditions. MU380 exhibited a stronger synergistic effect with GEM than clinical candidate SCH900776. MU380 alone or in combination with GEM significantly reduced spheroid size and increased apoptosis in all patient-derived xenograft 3D cultures, with a higher impact in DR models. Combined treatment induced premature mitosis from G1 phase resulting in the mitotic catastrophe as a prestage of apoptosis. Finally, treatment by MU380 alone, or in combination with GEM, significantly inhibited tumor growth of both PC339-DOC and PC346C-DOC xenograft models in mice. Taken together, our data suggest that metabolically robust and selective CHK1 inhibitor MU380 can bypass docetaxel resistance and improve the effectiveness of GEM in DR mCRPC models. This approach might allow for dose reduction of GEM and thereby minimize undesired toxicity and may represent a therapeutic option for patients with incurable DR mCRPC.},

note = {Place: United States},

keywords = {*Mitosis/drug effects, Animals, castration-resistant prostate cancer, Cell Death/drug effects, Cell Line, Cell Proliferation/drug effects, Checkpoint Kinase 1, Checkpoint Kinase 1/*antagonists & inhibitors/metabolism, Deoxycytidine/*analogs & derivatives/pharmacology, Docetaxel resistance, Docetaxel/*pharmacology, Drug resistance, gemcitabine, Humans, Male, Mice, mitotic catastrophe, MU380, Neoplasm/*drug effects, Piperidines/chemistry/*pharmacology, Prostatic Neoplasms/*pathology, Pyrazoles/chemistry/*pharmacology, Pyrimidines/chemistry/*pharmacology, S Phase/drug effects, SCID, Tumor, Xenograft Model Antitumor Assays},

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{kahounova_slug-expressing_2020,

title = {Slug-expressing mouse prostate epithelial cells have increased stem cell potential.},

author = {Zuzana Kahounová and Ján Remšík and Radek Fedr and Jan Bouchal and Alena Mičková and Eva Slabáková and Lucia Binó and Aleš Hampl and Karel Souček},

doi = {10.1016/j.scr.2020.101844},

issn = {1876-7753 1873-5061},

year  = {2020},

date = {2020-07-01},

journal = {Stem cell research},

volume = {46},

pages = {101844},

abstract = {Deciphering the properties of adult stem cells is crucial for understanding of their role in healthy tissue and in cancer progression as well. Both stem cells and cancer stem cells have shown association with epithelial-to-mesenchymal transition (EMT) in various tissue types. Aiming to investigate the epithelial and mesenchymal phenotypic traits in adult mouse prostate, we sorted subpopulations of basal prostate stem cells (mPSCs) and assessed the expression levels of EMT regulators and markers with custom-designed gene expression array. The population of mPSCs defined by a Lin(-)/Sca-1(+)CD49f(hi)/Trop-2(+) (LSC Trop-2(+)) surface phenotype was enriched in mesenchymal markers, especially EMT master regulator Slug, encoded by the Snai2 gene. To further dissect the role of Slug in mPSCs, we used transgenic Snai2(tm1.1Wbg) reporter mouse strain. Using this model, we confirmed the presence of mesenchymal traits and increase of organoid forming capacity in Slug(+) population of mPSCs. The Slug(+)-derived organoids comprised all prostate epithelial cell types - basal, luminal, and neuroendocrine. Collectively, these data uncover the important role of Slug expression in the physiology of mouse prostate stem cells.},

note = {Place: England},

keywords = {*Epithelial-Mesenchymal Transition, *Prostate, Animals, Cell Line, Cell Movement, Epithelial Cells, epithelial-to-mesenchymal transition, Male, Mice, Organoids, Prostate stem cells, Snai2/Slug, Snail Family Transcription Factors/genetics, stemness, Tumor},

pubstate = {published},

tppubtype = {article}

}

@article{vyhlidalova_gut_2020,

title = {Gut Microbial Catabolites of Tryptophan Are Ligands and Agonists of the Aryl Hydrocarbon Receptor: A Detailed Characterization.},

author = {Barbora Vyhlídalová and Kristýna Krasulová and Petra Pečinková and Adéla Marcalíková and Radim Vrzal and Lenka Zemánková and Jan Vančo and Zdeněk Trávníček and Jan Vondráček and Martina Karasová and Sridhar Mani and Zdeněk Dvořák},

doi = {10.3390/ijms21072614},

issn = {1422-0067},

year  = {2020},

date = {2020-04-01},

journal = {International journal of molecular sciences},

volume = {21},

number = {7},

abstract = {We examined the effects of gut microbial catabolites of tryptophan on the aryl hydrocarbon receptor (AhR). Using a reporter gene assay, we show that all studied catabolites are low-potency agonists of human AhR. The efficacy of catabolites differed substantially, comprising agonists with no or low (i3-propionate, i3-acetate, i3-lactate, i3-aldehyde), medium (i3-ethanol, i3-acrylate, skatole, tryptamine), and high (indole, i3-acetamide, i3-pyruvate) efficacies. We displayed ligand-selective antagonist activities by i3-pyruvate, i3-aldehyde, indole, skatole, and tryptamine. Ligand binding assay identified low affinity (skatole, i3-pyruvate, and i3-acetamide) and very low affinity (i3-acrylate, i3-ethanol, indole) ligands of the murine AhR. Indole, skatole, tryptamine, i3-pyruvate, i3-acrylate, and i3-acetamide induced CYP1A1 mRNA in intestinal LS180 and HT-29 cells, but not in the AhR-knockout HT-29 variant. We observed a similar CYP1A1 induction pattern in primary human hepatocytes. The most AhR-active catabolites (indole, skatole, tryptamine, i3-pyruvate, i3-acrylate, i3-acetamide) elicited nuclear translocation of the AhR, followed by a formation of AhR-ARNT heterodimer and enhanced binding of the AhR to the CYP1A1 gene promoter. Collectively, we comprehensively characterized the interactions of gut microbial tryptophan catabolites with the AhR, which may expand the current understanding of their potential roles in intestinal health and disease.},

note = {Place: Switzerland},

keywords = {*Gastrointestinal Microbiome/drug effects, Animals, Aryl hydrocarbon receptor, Aryl Hydrocarbon/*agonists/*metabolism, Basic Helix-Loop-Helix Transcription Factors/*agonists/*metabolism, Cell Line, Cytochrome P-450 CYP1A1/genetics, Gene Expression, Genes, Genetic, Humans, Indoles, Ligands, Metabolic Networks and Pathways, Mice, Microbiome, Promoter Regions, Protein Binding, Protein Multimerization, Receptors, Reporter, tryptophan, Tryptophan/*metabolism, Tumor},

pubstate = {published},

tppubtype = {article}

}

@article{boudny_novel_2019,

title = {Novel CHK1 inhibitor MU380 exhibits significant single-agent activity in TP53-mutated chronic lymphocytic leukemia cells.},

author = {Miroslav Boudny and Jana Zemanova and Prashant Khirsariya and Marek Borsky and Jan Verner and Jana Cerna and Alexandra Oltova and Vaclav Seda and Marek Mraz and Josef Jaros and Zuzana Jaskova and Michaela Spunarova and Yvona Brychtova and Karel Soucek and Stanislav Drapela and Marie Kasparkova and Jiri Mayer and Kamil Paruch and Martin Trbusek},

doi = {10.3324/haematol.2018.203430},

issn = {1592-8721 0390-6078},

year  = {2019},

date = {2019-12-01},

journal = {Haematologica},

volume = {104},

number = {12},

pages = {2443–2455},

abstract = {Introduction of small-molecule inhibitors of B-cell receptor signaling and BCL2 protein significantly improves therapeutic options in chronic lymphocytic leukemia. However, some patients suffer from adverse effects mandating treatment discontinuation, and cases with TP53 defects more frequently experience early progression of the disease. Development of alternative therapeutic approaches is, therefore, of critical importance. Here we report details of the anti-chronic lymphocytic leukemia single-agent activity of MU380, our recently identified potent, selective, and metabolically robust inhibitor of checkpoint kinase 1. We also describe a newly developed enantioselective synthesis of MU380, which allows preparation of gram quantities of the substance. Checkpoint kinase 1 is a master regulator of replication operating primarily in intra-S and G(2)/M cell cycle checkpoints. Initially tested in leukemia and lymphoma cell lines, MU380 significantly potentiated efficacy of gemcitabine, a clinically used inducer of replication stress. Moreover, MU380 manifested substantial single-agent activity in both TP53-wild type and TP53-mutated leukemia and lymphoma cell lines. In chronic lymphocytic leukemia-derived cell lines MEC-1, MEC-2 (both TP53-mut), and OSU-CLL (TP53-wt) the inhibitor impaired cell cycle progression and induced apoptosis. In primary clinical samples, MU380 used as a single-agent noticeably reduced the viability of unstimulated chronic lymphocytic leukemia cells as well as those induced to proliferate by anti-CD40/IL-4 stimuli. In both cases, effects were comparable in samples harboring p53 pathway dysfunction (TP53 mutations or ATM mutations) and TP53-wt/ATM-wt cells. Lastly, MU380 also exhibited significant in vivo activity in a xenotransplant mouse model (immunodeficient strain NOD-scid IL2Rγ(null) ) where it efficiently suppressed growth of subcutaneous tumors generated from MEC-1 cells.},

note = {Place: Italy},

keywords = {*Drug Synergism, *Mutation, Animals, Antimetabolites, Antineoplastic/pharmacology, Apoptosis, B-Cell/*drug therapy/genetics/pathology, Biomarkers, Cell Cycle, Cell Proliferation, Checkpoint Kinase 1/*antagonists & inhibitors, Chronic, Cultured, Deoxycytidine/analogs & derivatives/pharmacology, Drug resistance, Female, gemcitabine, Gene Expression Regulation, Humans, Inbred NOD, Leukemia, Lymphocytic, Mice, Neoplasm/drug effects, Neoplastic/*drug effects, Piperidines/*pharmacology, Protein Kinase Inhibitors/pharmacology, Pyrazoles/*pharmacology, Pyrimidines/*pharmacology, SCID, Tumor Cells, Tumor Suppressor Protein p53/*genetics, Tumor/genetics, Xenograft Model Antitumor Assays},

pubstate = {published},

tppubtype = {article}

}

@article{jirik_blind_2019,

title = {Blind deconvolution estimation of an arterial input function for small animal DCE-MRI.},

author = {Radovan Jiřík and Torfinn Taxt and Ondřej Macíček and Michal Bartoš and Jiří Kratochvíla and Karel Souček and Eva Dražanová and Lucie Krátká and Aleš Hampl and Zenon Jr Starčuk},

doi = {10.1016/j.mri.2019.05.024},

issn = {1873-5894 0730-725X},

year  = {2019},

date = {2019-10-01},

journal = {Magnetic resonance imaging},

volume = {62},

pages = {46–56},

abstract = {PURPOSE: One of the main obstacles for reliable quantitative dynamic contrast-enhanced (DCE) MRI is the need for accurate knowledge of the arterial input function (AIF). This is a special challenge for preclinical small animal applications where it is very difficult to measure the AIF without partial volume and flow artifacts. Furthermore, using advanced pharmacokinetic models (allowing estimation of blood flow and permeability-surface area product in addition to the classical perfusion parameters) poses stricter requirements on the accuracy and precision of AIF estimation. This paper addresses small animal DCE-MRI with advanced pharmacokinetic models and presents a method for estimation of the AIF based on blind deconvolution. METHODS: A parametric AIF model designed for small animal physiology and use of advanced pharmacokinetic models is proposed. The parameters of the AIF are estimated using multichannel blind deconvolution. RESULTS: Evaluation on simulated data show that for realistic signal to noise ratios blind deconvolution AIF estimation leads to comparable results as the use of the true AIF. Evaluation on real data based on DCE-MRI with two contrast agents of different molecular weights showed a consistence with the known effects of the molecular weight. CONCLUSION: Multi-channel blind deconvolution using the proposed AIF model specific for small animal DCE-MRI provides reliable perfusion parameter estimates under realistic signal to noise conditions.},

note = {Place: Netherlands},

keywords = {*Magnetic Resonance Imaging, Algorithms, Animals, Arterial input function, Arteries/*diagnostic imaging, Blind deconvolution, Computer Simulation, Computer-Assisted/*methods, Contrast Media/*pharmacokinetics, DCE-MRI, Humans, Image Processing, Inbred BALB C, Mice, Necrosis/pathology, Perfusion, Pharmacokinetics, Regression Analysis, Reproducibility of Results, Signal-To-Noise Ratio},

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}

}

@article{simek_lc-msms_2019,

title = {LC-MS/MS study of in vivo fate of hyaluronan polymeric micelles carrying doxorubicin.},

author = {Matěj Šimek and Martina Hermannová and Daniela Šmejkalová and Tereza Foglová and Karel Souček and Lucia Binó and Vladimír Velebný},

doi = {10.1016/j.carbpol.2018.12.104},

issn = {1879-1344 0144-8617},

year  = {2019},

date = {2019-04-01},

journal = {Carbohydrate polymers},

volume = {209},

pages = {181–189},

abstract = {A better understanding of in vivo behavior of nanocarriers is necessary for further improvement in their development. Here we present a novel approach, where both the matrix and the drug can be analyzed by LCMS/MS after one sample handling. The developed method was applied for the comparison of pharmacokinetic profile of free and encapsulated doxorubicin (DOX) in oleyl hyaluronan (HA-C18:1) polymeric micelles. The results indicated that nanocarriers were rapidly dissociated upon in vivo administration. Despite this fact, the administration of encapsulated DOX led to its longer circulation time and enhanced tumor targeting. This effect was not observed injecting blank HA-C18:1 micelles followed by unencapsulated DOX. Biodistribution studies and molecular weight estimation of the carrier matrix indicated relatively high stability of HA-C18:1 ester bond in bloodstream and complete elimination of the derivative within 72 h. The proposed methodology provides a novel strategy to elucidate the pharmacokinetic behavior of polysaccharide-based drug delivery systems.},

note = {Place: England},

keywords = {*Micelles, Animals, Biodistribution, Chromatography, Doxorubicin, Doxorubicin/*chemistry/pharmacokinetics, Drug Carriers/*chemistry, Drug Liberation, Female, Hyaluronan, Hyaluronic Acid/*chemistry, Liquid, Mice, Molecular Weight, Pharmacokinetics, Polymeric micelles, Tandem Mass Spectrometry, Tissue Distribution},

pubstate = {published},

tppubtype = {article}

}

@article{remsik_trop-2_2018,

title = {Trop-2 plasticity is controlled by epithelial-to-mesenchymal transition.},

author = {Ján Remšík and Lucia Binó and Zuzana Kahounová and Gvantsa Kharaishvili and Šárka Šimecková and Radek Fedr and Tereza Kucírková and Sára Lenárt and Ximena Maria Muresan and Eva Slabáková and Lucia Knopfová and Jan Bouchal and Milan Král and Petr Beneš and Karel Soucek},

doi = {10.1093/carcin/bgy095},

issn = {1460-2180 0143-3334},

year  = {2018},

date = {2018-12-01},

journal = {Carcinogenesis},

volume = {39},

number = {11},

pages = {1411–1418},

abstract = {The cell surface glycoprotein Trop-2 is commonly overexpressed in carcinomas and represents an exceptional antigen for targeted therapy. Here, we provide evidence that surface Trop-2 expression is functionally connected with an epithelial phenotype in breast and prostate cell lines and in patient tumor samples. We further show that Trop-2 expression is suppressed epigenetically or through the action of epithelial-to-mesenchymal transition transcription factors and that deregulation of Trop-2 expression is linked with cancer progression and poor patient prognosis. Moreover, our data suggest that the cancer plasticity-driven intratumoral heterogeneity in Trop-2 expression may significantly contribute to response and resistance to therapies targeting Trop-2-expressing cells.},

note = {Place: England},

keywords = {Animals, Antigens, Breast Neoplasms/mortality/*pathology, Cadherins/biosynthesis, Carcinoma/*pathology, CD/biosynthesis, Cell Adhesion Molecules/genetics/*metabolism, Cell Line, Disease Progression, DNA Methylation/genetics, Epithelial Cells/*metabolism, Epithelial-Mesenchymal Transition/physiology, Female, Humans, Inbred BALB C, Male, Mice, Neoplasm/genetics/*metabolism, Prostatic Neoplasms/mortality/*pathology, Tumor, Xenograft Model Antitumor Assays},

pubstate = {published},

tppubtype = {article}

}

@article{strapacova_relative_2018,

title = {Relative effective potencies of dioxin-like compounds in rodent and human lung cell models.},

author = {Simona Strapáčová and Petra Brenerová and Pavel Krčmář and Patrik Andersson and Karin I. Ede and Majorie B. M. Duursen and Martin Berg and Jan Vondráček and Miroslav Machala},

doi = {10.1016/j.tox.2018.05.004},

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

year  = {2018},

date = {2018-07-01},

journal = {Toxicology},

volume = {404-405},

pages = {33–41},

abstract = {Toxicity of dioxin-like compounds (DLCs), such as polychlorinated dibenzo-p-dioxins, dibenzofurans and biphenyls, is largely mediated via aryl hydrocarbon receptor (AhR) activation. AhR-mediated gene expression can be tissue-specific; however, the inducibility of AhR in the lungs, a major target of DLCs, remains poorly characterized. In this study, we developed relative effective potencies (REPs) for a series of DLCs in both rodent (MLE-12, RLE-6TN) and human (A549, BEAS-2B) lung and bronchial epithelial cell models, using expression of both canonical (CYP1A1, CYP1B1) and less well characterized (TIPARP, AHRR, ALDH3A1) AhR target genes. The use of rat, murine and human cell lines allowed us to determine both species-specific differences in sensitivity of responses to DLCs in lung cellular models and deviations from established WHO toxic equivalency factor values (TEF) values. Finally, expression of selected AhR target genes was determined in vivo, using lung tissues of female rats exposed to a single oral dose of DLCs and compared with the obtained in vitro data. All cell models were highly sensitive to DLCs, with murine MLE-12 cells being the most sensitive and human A549 cells being the least sensitive. Interestingly, we observed that four AhR target genes were more sensitive than CYP1A1 in lung cell models (CYP1B1, AHRR, TIPARP and/or ALDH3A1). We found some deviations, with strikingly low REPs for polychlorinated biphenyls PCBs 105, 167, 169 and 189 in rat RLE-6TN cells-derived REPs for a series of 20 DLCs evaluated in this study, as compared with WHO TEF values. For other DLCs, including PCBs 126, 118 and 156, REPs were generally in good accordance with WHO TEF values. This conclusion was supported by in vivo data obtained in rat lung tissue. However, we found that human lung REPs for 2,3,4,7,8-pentachlorodibenzofuran and PCB 126 were much lower than the respective rat lung REPs. Furthermore, PCBs 118 and 156 were almost inactive in these human cells. Our observations may have consequences for risk assessment. Given the differences observed between rat and human data sets, development of human-specific REP/TEFs, and the use of CYP1B1, AHRR, TIPARP and/or ALDH3A1 mRNA inducibility as sensitive endpoints, are recommended for assessment of relative effective potencies of DLCs.},

note = {Place: Ireland},

keywords = {A549 Cells, Acute/methods, AhR, Animals, Dioxin-like compounds, Dioxins/*toxicity, Dose-Response Relationship, Drug, Endogenous target genes, Female, Humans, Lung epithelial cells, Lung/*drug effects/metabolism/*pathology, Mice, Rats, Relative effective potencies, Rodentia, Species Specificity, Sprague-Dawley, Toxicity Tests},

pubstate = {published},

tppubtype = {article}

}

@article{vargova_hypericin_2018,

title = {Hypericin affects cancer side populations via competitive inhibition of BCRP.},

author = {Jana Vargová and Jaromír Mikeš and Rastislav Jendželovský and Lucia Mikešová and Barbora Kuchárová and Ľubomír Čulka and Radek Fedr and Ján Remšík and Karel Souček and Alois Kozubík and Peter Fedoročko},

doi = {10.1016/j.biopha.2018.01.074},

issn = {1950-6007 0753-3322},

year  = {2018},

date = {2018-03-01},

journal = {Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie},

volume = {99},

pages = {511–522},

abstract = {OBJECTIVE: Cancer stem-like cells (CSLCs) are considered a root of tumorigenicity and resistance. However, their identification remains challenging. The use of the side population (SP) assay as a credible marker of CSLCs remains controversial. The SP assay relies on the elevated activity of ABC transporters that, in turn, can be modulated by hypericin (HYP), a photosensitizer and bioactive compound of St. John's Wort (Hypericum perforatum), a popular over-the-counter antidepressant. Here we aimed to comprehensively characterize the SP phenotype of cancer cells and to determine the impact of HYP on these cells. METHODS: Flow cytometry and sorting-based assays were employed, including CD24-, CD44-, CD133-, and ALDH-positivity, clonogenicity, 3D-forming ability, ABC transporter expression and activity, and intracellular accumulation of HYP/Hoechst 33342. The tumorigenic ability of SP, nonSP, and HYP-treated cells was verified by xenotransplantation into immunodeficient mice. RESULTS: The SP phenotype was associated with elevated expression of several investigated transporters and more intensive growth in non-adherent conditions but not with higher clonogenicity, tumorigenicity or ALDH-positivity. Despite stimulated BCRP level and MRP1 activity, HYP reversibly decreased the SP proportion, presumably via competitive inhibition of BCRP. HYP-selected SP cells acquired additional traits of resistance and extensively eliminated HYP. CONCLUSIONS: Our results suggest that SP is not an unequivocal CSLC-marker. However, SP could play an important role in modulating HYP-treatment and serve as a negative predictive tool for HYP-based therapies. Moreover, the use of supplements containing HYP by cancer patients should be carefully considered, due to its proposed effect on drug efflux and complex impact on tumor cells, which have not yet been sufficiently characterized.},

note = {Place: France},

keywords = {ABC transporters, Aldehyde Dehydrogenase/metabolism, Animals, Anthracenes, ATP Binding Cassette Transporter, Biomarkers, Cancer stem-like cells, Carcinogenesis/drug effects/metabolism/pathology, Cell Line, Cellular/drug effects/metabolism/pathology, Clone Cells, Drug resistance, Humans, Hypericin, Member 1/metabolism, Member 2/*metabolism, Mice, Neoplasm Proteins/*metabolism, Neoplasms/*metabolism/*pathology, Neoplastic Stem Cells/drug effects/metabolism/pathology, Perylene/*analogs & derivatives/pharmacology, Phenotype, SCID, Side population, Side-Population Cells/drug effects/*pathology, Spheroids, St. John’s wort, Subfamily B, Subfamily G, Substrate Specificity/drug effects, Survival Analysis, Tumor, Tumor/metabolism},

pubstate = {published},

tppubtype = {article}

}

@article{paculova_brca1_2017,

title = {BRCA1 or CDK12 loss sensitizes cells to CHK1 inhibitors.},

author = {Hana Paculová and Juraj Kramara and Šárka Šimečková and Radek Fedr and Karel Souček and Ondřej Hylse and Kamil Paruch and Marek Svoboda and Martin Mistrík and Jiří Kohoutek},

doi = {10.1177/1010428317727479},

issn = {1423-0380 1010-4283},

year  = {2017},

date = {2017-10-01},

journal = {Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine},

volume = {39},

number = {10},

pages = {1010428317727479},

abstract = {A broad spectrum of tumors develop resistance to classic chemotherapy, necessitating the discovery of new therapies. One successful strategy exploits the synthetic lethality between poly(ADP-ribose) polymerase 1/2 proteins and DNA damage response genes, including BRCA1, a factor involved in homologous recombination-mediated DNA repair, and CDK12, a transcriptional kinase known to regulate the expression of DDR genes. CHK1 inhibitors have been shown to enhance the anti-cancer effect of DNA-damaging compounds. Since loss of BRCA1 increases replication stress and leads to DNA damage, we tested a hypothesis that CDK12- or BRCA1-depleted cells rely extensively on S-phase-related CHK1 functions for survival. The silencing of BRCA1 or CDK12 sensitized tumor cells to CHK1 inhibitors in vitro and in vivo. BRCA1 downregulation combined with CHK1 inhibition induced excessive amounts of DNA damage, resulting in an inability to complete the S-phase. Therefore, we suggest CHK1 inhibition as a strategy for targeting BRCA1- or CDK12-deficient tumors.},

note = {Place: Netherlands},

keywords = {Animals, BRCA1, BRCA1 Protein/antagonists & inhibitors/*genetics, CDK12, Checkpoint Kinase 1/*genetics, CHK1 inhibitor, Colorectal Neoplasms/drug therapy/*genetics/pathology, Cyclin-Dependent Kinases/antagonists & inhibitors/*genetics, DNA damage response, DNA Damage/drug effects, Drug resistance, Gene Expression Regulation, Gene Silencing, HCT116 Cells, Humans, Mice, Neoplasm/genetics, Neoplastic/drug effects, Poly (ADP-Ribose) Polymerase-1/genetics, Pyrazoles/administration & dosage, Pyrimidines/administration & dosage, Transcription, Xenograft Model Antitumor Assays},

pubstate = {published},

tppubtype = {article}

}

@article{samadder_synthesis_2017,

title = {Synthesis and Profiling of a Novel Potent Selective Inhibitor of CHK1 Kinase Possessing Unusual N-trifluoromethylpyrazole Pharmacophore Resistant to Metabolic N-dealkylation.},

author = {Pounami Samadder and Tereza Suchánková and Ondřej Hylse and Prashant Khirsariya and Fedor Nikulenkov and Stanislav Drápela and Nicol Straková and Petr Vaňhara and Kateřina Vašíčková and Hana Kolářová and Lucia Binó and Miroslava Bittová and Petra Ovesná and Peter Kollár and Radek Fedr and Milan Ešner and Josef Jaroš and Aleš Hampl and Lumír Krejčí and Kamil Paruch and Karel Souček},

doi = {10.1158/1535-7163.MCT-17-0018},

issn = {1538-8514 1535-7163},

year  = {2017},

date = {2017-09-01},

journal = {Molecular cancer therapeutics},

volume = {16},

number = {9},

pages = {1831–1842},

abstract = {Checkpoint-mediated dependency of tumor cells can be deployed to selectively kill them without substantial toxicity to normal cells. Specifically, loss of CHK1, a serine threonine kinase involved in the surveillance of the G(2)-M checkpoint in the presence of replication stress inflicted by DNA-damaging drugs, has been reported to dramatically influence the viability of tumor cells. CHK1's pivotal role in maintaining genomic stability offers attractive opportunity for increasing the selectivity, effectivity, and reduced toxicity of chemotherapy. Some recently identified CHK1 inhibitors entered clinical trials in combination with DNA antimetabolites. Herein, we report synthesis and profiling of MU380, a nontrivial analogue of clinically profiled compound SCH900776 possessing the highly unusual N-trifluoromethylpyrazole motif, which was envisioned not to undergo metabolic oxidative dealkylation and thereby provide greater robustness to the compound. MU380 is a selective and potent inhibitor of CHK1 which sensitizes a variety of tumor cell lines to hydroxyurea or gemcitabine up to 10 times. MU380 shows extended inhibitory effects in cells, and unlike SCH900776, does not undergo in vivo N-dealkylation to the significantly less selective metabolite. Compared with SCH900776, MU380 in combination with GEM causes higher accumulation of DNA damage in tumor cells and subsequent enhanced cell death, and is more efficacious in the A2780 xenograft mouse model. Overall, MU380 represents a novel state-of-the-art CHK1 inhibitor with high potency, selectivity, and improved metabolic robustness to oxidative N-dealkylation. Mol Cancer Ther; 16(9); 1831-42. ©2017 AACR.},

note = {Place: United States},

keywords = {Animal, Animals, Antineoplastic Agents/*chemical synthesis/*pharmacology, Apoptosis/drug effects, Biomarkers, Cell Cycle Checkpoints/drug effects, Cell Cycle/drug effects, Cell Line, Checkpoint Kinase 1/*antagonists & inhibitors, Dealkylation/drug effects, Disease Models, Dose-Response Relationship, Drug, Drug resistance, Humans, Methylation, Mice, Molecular Structure, Neoplasm/*drug effects, Protein Kinase Inhibitors/*chemical synthesis/*pharmacology, Pyrazoles/pharmacology, Pyrimidines/pharmacology, Tumor, Xenograft Model Antitumor Assays},

pubstate = {published},

tppubtype = {article}

}

@article{kremserova_lung_2016,

title = {Lung Neutrophilia in Myeloperoxidase Deficient Mice during the Course of Acute Pulmonary Inflammation.},

author = {Silvie Kremserova and Tomas Perecko and Karel Soucek and Anna Klinke and Stephan Baldus and Jason P. Eiserich and Lukas Kubala},

doi = {10.1155/2016/5219056},

issn = {1942-0994 1942-0900},

year  = {2016},

date = {2016-01-01},

journal = {Oxidative medicine and cellular longevity},

volume = {2016},

pages = {5219056},

abstract = {Systemic inflammation accompanying diseases such as sepsis affects primarily lungs and induces their failure. This remains the most common cause of sepsis induced mortality. While neutrophils play a key role in pulmonary failure, the mechanisms remain incompletely characterized. We report that myeloperoxidase (MPO), abundant enzyme in neutrophil granules, modulates the course of acute pulmonary inflammatory responses induced by intranasal application of lipopolysaccharide. MPO deficient mice had significantly increased numbers of airway infiltrated neutrophils compared to wild-type mice during the whole course of lung inflammation. This was accompanied by higher levels of RANTES in bronchoalveolar lavage fluid from the MPO deficient mice. Other markers of lung injury and inflammation, which contribute to recruitment of neutrophils into the inflamed lungs, including total protein and other selected proinflammatory cytokines did not significantly differ in bronchoalveolar lavage fluid from the wild-type and the MPO deficient mice. Interestingly, MPO deficient neutrophils revealed a decreased rate of cell death characterized by phosphatidylserine surface expression. Collectively, the importance of MPO in regulation of pulmonary inflammation, independent of its putative microbicidal functions, can be potentially linked to MPO ability to modulate the life span of neutrophils and to affect accumulation of chemotactic factors at the inflammatory site.},

note = {Place: United States},

keywords = {Acute Disease, Acute Lung Injury/complications/genetics, Animals, Inborn Errors/*complications, Inbred C57BL, Knockout, Leukocyte Disorders/*complications/*genetics, Lipopolysaccharides, Male, metabolism, Mice, Neutrophils/*pathology, Peroxidase/deficiency/genetics, Pneumonia/chemically induced/*complications/genetics},

pubstate = {published},

tppubtype = {article}

}

@article{slabakova_opposite_2015,

title = {Opposite regulation of MDM2 and MDMX expression in acquisition of mesenchymal phenotype in benign and cancer cells.},

author = {Eva Slabáková and Gvantsa Kharaishvili and Monika Smějová and Zuzana Pernicová and Tereza Suchánková and Ján Remšík and Stanislav Lerch and Nicol Straková and Jan Bouchal and Milan Král and Zoran Culig and Alois Kozubík and Karel Souček},

doi = {10.18632/oncotarget.5392},

issn = {1949-2553},

year  = {2015},

date = {2015-11-01},

journal = {Oncotarget},

volume = {6},

number = {34},

pages = {36156–36171},

abstract = {Plasticity of cancer cells, manifested by transitions between epithelial and mesenchymal phenotypes, represents a challenging issue in the treatment of neoplasias. Both epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial transition (MET) are implicated in the processes of metastasis formation and acquisition of stem cell-like properties. Mouse double minute (MDM) 2 and MDMX are important players in cancer progression, as they act as regulators of p53, but their function in EMT and metastasis may be contradictory. Here, we show that the EMT phenotype in multiple cellular models and in clinical prostate and breast cancer samples is associated with a decrease in MDM2 and increase in MDMX expression. Modulation of EMT-accompanying changes in MDM2 expression in benign and transformed prostate epithelial cells influences their migration capacity and sensitivity to docetaxel. Analysis of putative mechanisms of MDM2 expression control demonstrates that in the context of defective p53 function, MDM2 expression is regulated by EMT-inducing transcription factors Slug and Twist. These results provide an alternative context-specific role of MDM2 in EMT, cell migration, metastasis, and therapy resistance.},

note = {Place: United States},

keywords = {Animals, Breast Neoplasms/genetics/*metabolism/pathology, Cell Cycle Proteins, Cell Line, Epithelial-Mesenchymal Transition, Epithelial-Mesenchymal Transition/*physiology, Female, Heterografts, Humans, Male, MDM2/MDMX, Mice, Nuclear Proteins/*biosynthesis, Nude, Phenotype, prostate/breast cancer, Prostatic Neoplasms/genetics/*metabolism/pathology, Proto-Oncogene Proteins c-mdm2/*biosynthesis, Proto-Oncogene Proteins/*biosynthesis, Snai2/Slug, Transfection, Tumor, TWIST},

pubstate = {published},

tppubtype = {article}

}

@article{kratochvilova_tumor_2015,

title = {Tumor suppressor candidate 3 (TUSC3) prevents the epithelial-to-mesenchymal transition and inhibits tumor growth by modulating the endoplasmic reticulum stress response in ovarian cancer cells.},

author = {Kateřina Kratochvílová and Peter Horak and Milan Ešner and Karel Souček and Dietmar Pils and Mariam Anees and Erwin Tomasich and František Dráfi and Veronika Jurtíková and Aleš Hampl and Michael Krainer and Petr Vaňhara},

doi = {10.1002/ijc.29502},

issn = {1097-0215 0020-7136},

year  = {2015},

date = {2015-09-01},

journal = {International journal of cancer},

volume = {137},

number = {6},

pages = {1330–1340},

abstract = {Ovarian cancer is one of the most common malignancies in women and contributes greatly to cancer-related deaths. Tumor suppressor candidate 3 (TUSC3) is a putative tumor suppressor gene located at chromosomal region 8p22, which is often lost in epithelial cancers. Epigenetic silencing of TUSC3 has been associated with poor prognosis, and hypermethylation of its promoter provides an independent biomarker of overall and disease-free survival in ovarian cancer patients. TUSC3 is localized to the endoplasmic reticulum in an oligosaccharyl tranferase complex responsible for the N-glycosylation of proteins. However, the precise molecular role of TUSC3 in ovarian cancer remains unclear. In this study, we establish TUSC3 as a novel ovarian cancer tumor suppressor using a xenograft mouse model and demonstrate that loss of TUSC3 alters the molecular response to endoplasmic reticulum stress and induces hallmarks of the epithelial-to-mesenchymal transition in ovarian cancer cells. In summary, we have confirmed the tumor-suppressive function of TUSC3 and identified the possible mechanism driving TUSC3-deficient ovarian cancer cells toward a malignant phenotype.},

note = {Place: United States},

keywords = {Animals, Cell Line, Endoplasmic Reticulum Stress, Endoplasmic Reticulum Stress/*genetics, Epithelial-Mesenchymal Transition/*genetics, epithelial-to-mesenchymal transition, Female, Genes, Heterografts, Humans, Inbred NOD, Membrane Proteins/*genetics, Mice, N33, ovarian cancer, Ovarian Neoplasms/*genetics, SCID, Tumor, Tumor Suppressor, Tumor Suppressor Proteins/*genetics, Tumor Suppressor/physiology, TUSC3},

pubstate = {published},

tppubtype = {article}

}

@article{smerdova_upregulation_2014,

title = {Upregulation of CYP1B1 expression by inflammatory cytokines is mediated by the p38 MAP kinase signal transduction pathway.},

author = {Lenka Smerdová and Jana Šmerdová and Markéta Kabátková and Jiří Kohoutek and Dalibor Blažek and Miroslav Machala and Jan Vondráček},

doi = {10.1093/carcin/bgu190},

issn = {1460-2180 0143-3334},

year  = {2014},

date = {2014-11-01},

journal = {Carcinogenesis},

volume = {35},

number = {11},

pages = {2534–2543},

abstract = {Cytochrome P450 1B1 (CYP1B1) is an enzyme that has a unique tumor-specific pattern of expression and is capable of bioactivating a wide range of carcinogenic compounds. We have reported previously that coordinated upregulation of CYP1B1 by inflammatory cytokines, such as tumor necrosis factor-α (TNF-α) and the aryl hydrocarbon receptor ligands, may increase bioactivation of promutagens, such as benzo[a]pyrene (BaP) in epithelial cells. Here, we extend those studies by describing a novel mechanism participating in the regulation of CYP1B1 expression, which involves activation of the p38 mitogen-activated protein kinase (p38) and mitogen- and stress-activated protein kinase 1 (MSK1). Using inhibitors of p38 and MSKs, as well as mouse embryonic cells derived from p38α-deficient and MSK1/2 double knockout mice, we show here that TNF-α potentiates CYP1B1 upregulation via the p38/MSK1 kinase cascade. Effects of this inflammatory cytokine on CYP1B1 expression further involve the positive transcription elongation factor b (P-TEFb). The inhibition of the P-TEFb subunit, cyclin-dependent kinase 9 (CDK9), which phosphorylates RNA polymerase II (RNAPII), prevented the enhanced CYP1B1 induction by a combination of BaP and inflammatory cytokine. Furthermore, using chromatin immunoprecipitation assays, we found that cotreatment of epithelial cells with TNF-α and BaP resulted in enhanced recruitment of both CDK9 and RNAPII to the Cyp1b1 gene promoter. Overall, these results have implications concerning the contribution of inflammatory factors to carcinogenesis, since enhanced CYP1B1 induction during inflammation may alter metabolism of exogenous carcinogens, as well as endogenous CYP1B1 substrates playing role in tumor development.},

note = {Place: England},

keywords = {Animals, Carcinogenesis/drug effects/*genetics, Carcinogens/toxicity, Cyclin-Dependent Kinase 9/genetics, Cytochrome P-450 CYP1B1/*biosynthesis/genetics, Cytokines/metabolism, Gene Expression Regulation, Humans, Mice, Neoplasms/chemically induced/*genetics/pathology, Neoplastic/drug effects, p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors/*genetics/metabolism, Positive Transcriptional Elongation Factor B/genetics, RNA Polymerase II/genetics, Signal Transduction/drug effects, Tumor Necrosis Factor-alpha/metabolism},

pubstate = {published},

tppubtype = {article}

}

@article{ghorbanzadeh_vitro_2014,

title = {In vitro and in silico derived relative effect potencies of ah-receptor-mediated effects by PCDD/Fs and PCBs in rat, mouse, and guinea pig CALUX cell lines.},

author = {Mehdi Ghorbanzadeh and Karin I. Ede and Malin Larsson and Majorie B. M. Duursen and Lorenz Poellinger and Sandra Lücke-Johansson and Miroslav Machala and Kateřina Pěnčíková and Jan Vondráček and Martin Berg and Michael S. Denison and Tine Ringsted and Patrik L. Andersson},

doi = {10.1021/tx5001255},

issn = {1520-5010 0893-228X},

year  = {2014},

date = {2014-07-01},

journal = {Chemical research in toxicology},

volume = {27},

number = {7},

pages = {1120–1132},

abstract = {For a better understanding of species-specific relative effect potencies (REPs), responses of dioxin-like compounds (DLCs) were assessed. REPs were calculated using chemical-activated luciferase gene expression assays (CALUX) derived from guinea pig, rat, and mouse cell lines. Almost all 20 congeners tested in the rodent cell lines were partial agonists and less efficacious than 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). For this reason, REPs were calculated for each congener using concentrations at which 20% of the maximal TCDD response was reached (REP20TCDD). REP20TCDD values obtained for PCDD/Fs were comparable with their toxic equivalency factors assigned by the World Health Organization (WHO-TEF), while those for PCBs were in general lower than the WHO-TEF values. Moreover, the guinea pig cell line was the most sensitive as indicated by the 20% effect concentrations of TCDD of 1.5, 5.6, and 11.0 pM for guinea pig, rat, and mouse cells, respectively. A similar response pattern was observed using multivariate statistical analysis between the three CALUX assays and the WHO-TEFs. The mouse assay showed minor deviation due to higher relative induction potential for 2,3,7,8-tetrachlorodibenzofuran and 2,3,4,6,7,8-hexachlorodibenzofuran and lower for 1,2,3,4,6,7,8-heptachlorodibenzofuran and 3,3',4,4',5-pentachlorobiphenyl (PCB126). 2,3,7,8-Tetrachlorodibenzofuran was more than two times more potent in the mouse assay as compared with that of rat and guinea pig cells, while measured REP20TCDD for PCB126 was lower in mouse cells (0.05) as compared with that of the guinea pig (0.2) and rat (0.07). In order to provide REP20TCDD values for all WHO-TEF assigned compounds, quantitative structure-activity relationship (QSAR) models were developed. The QSAR models showed that specific electronic properties and molecular surface characteristics play important roles in the AhR-mediated response. In silico derived REP20TCDD values were generally consistent with the WHO-TEFs with a few exceptions. The QSAR models indicated that, e.g., 1,2,3,7,8-pentachlorodibenzofuran and 1,2,3,7,8,9-hexachlorodibenzofuran were more potent than given by their assigned WHO-TEF values, and the non-ortho PCB 81 was predicted, based on the guinea-pig model, to be 1 order of magnitude above its WHO-TEF value. By combining in vitro and in silico approaches, REPs were established for all WHO-TEF assigned compounds (except OCDD), which will provide future guidance in testing AhR-mediated responses of DLCs and to increase our understanding of species variation in AhR-mediated effects.},

note = {Place: United States},

keywords = {Animals, Aryl Hydrocarbon/agonists/*metabolism, Benzofurans/*pharmacology, Biological, Biological Assay, Cell Line, Computer Simulation, Dibenzofurans, Dose-Response Relationship, Drug, Guinea Pigs, Luciferases/metabolism, Mice, Models, Polychlorinated, Polychlorinated Biphenyls/*pharmacology, Polychlorinated Dibenzodioxins/*analogs & derivatives/pharmacology, Quantitative Structure-Activity Relationship, Rats, Receptors, Tumor},

pubstate = {published},

tppubtype = {article}

}

@article{jirik_blind_2014,

title = {Blind deconvolution in dynamic contrast-enhanced MRI and ultrasound.},

author = {Radovan Jiřík and Karel Souček and Martin Mézl and Michal Bartoš and Eva Dražanová and František Dráfi and Lucie Grossová and Jiří Kratochvíla and Ondřej Macíček and Kim Nylund and Aleš Hampl and Odd Helge Gilja and Torfinn Taxt and Zenon Jr Starčuk},

doi = {10.1109/EMBC.2014.6944569},

issn = {2694-0604 2375-7477},

year  = {2014},

date = {2014-01-01},

journal = {Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Annual International Conference},

volume = {2014},

pages = {4276–4279},

abstract = {This paper is focused on quantitative perfusion analysis using MRI and ultrasound. In both MRI and ultrasound, most approaches allow estimation of rate constants (Ktrans, kep for MRI) and indices (AUC, TTP) that are only related to the physiological perfusion parameters of a tissue (e.g. blood flow, vessel permeability) but do not allow their absolute quantification. Recent methods for quantification of these physiological perfusion parameters are shortly reviewed. The main problem of these methods is estimation of the arterial input function (AIF). This paper summarizes and extends the current blind-deconvolution approaches to AIF estimation. The feasibility of these methods is shown on a small preclinical study using both MRI and ultrasound.},

note = {Place: United States},

keywords = {Animals, Cell Line, Contrast Media/*pharmacokinetics, Experimental/diagnostic imaging/metabolism, Gadolinium DTPA/*pharmacokinetics, Humans, Inbred BALB C, Magnetic Resonance Imaging/methods, Mice, Neoplasm Transplantation, Neoplasms, Tissue Distribution, Tumor, Ultrasonography},

pubstate = {published},

tppubtype = {article}

}

@article{knopfova_c-myb_2012,

title = {c-Myb regulates matrix metalloproteinases 1/9, and cathepsin D: implications for matrix-dependent breast cancer cell invasion and metastasis.},

author = {Lucia Knopfová and Petr Beneš and Lucie Pekarčíková and Markéta Hermanová and Michal Masařík and Zuzana Pernicová and Karel Souček and Jan Smarda},

doi = {10.1186/1476-4598-11-15},

issn = {1476-4598},

year  = {2012},

date = {2012-03-01},

journal = {Molecular cancer},

volume = {11},

pages = {15},

abstract = {BACKGROUND: The c-Myb transcription factor is essential for the maintenance of stem-progenitor cells in bone marrow, colon epithelia, and neurogenic niches. c-Myb malfunction contributes to several types of malignancies including breast cancer. However, the function of c-Myb in the metastatic spread of breast tumors remains unexplored. In this study, we report a novel role of c-Myb in the control of specific proteases that regulate the matrix-dependent invasion of breast cancer cells. RESULTS: Ectopically expressed c-Myb enhanced migration and ability of human MDA-MB-231 and mouse 4T1 mammary cancer cells to invade Matrigel but not the collagen I matrix in vitro. c-Myb strongly increased the expression/activity of cathepsin D and matrix metalloproteinase (MMP) 9 and significantly downregulated MMP1. The gene coding for cathepsin D was suggested as the c-Myb-responsive gene and downstream effector of the migration-promoting function of c-Myb. Finally, we demonstrated that c-Myb delayed the growth of mammary tumors in BALB/c mice and affected the metastatic potential of breast cancer cells in an organ-specific manner. CONCLUSIONS: This study identified c-Myb as a matrix-dependent regulator of invasive behavior of breast cancer cells.},

note = {Place: England},

keywords = {Animals, Breast Neoplasms/genetics/*metabolism, Cathepsin D/genetics/*metabolism, Cell Line, Cell Movement/genetics/physiology, Electrophoresis, Female, Humans, Immunoblotting, Inbred BALB C, Matrix Metalloproteinase 1/genetics/*metabolism, Matrix Metalloproteinase 9/genetics/*metabolism, Mice, Neoplasm Metastasis/genetics/physiopathology, Polyacrylamide Gel, Proto-Oncogene Proteins c-myb/genetics/*metabolism, Real-Time Polymerase Chain Reaction, RNA, Small Interfering, Tumor},

pubstate = {published},

tppubtype = {article}

}

@article{uhlirova_suv39h-_2010,

title = {SUV39h- and A-type lamin-dependent telomere nuclear rearrangement.},

author = {Radka Uhlírová and Andrea Harnicarová Horáková and Gabriela Galiová and Sona Legartová and Pavel Matula and Miloslava Fojtová and Miroslav Varecha and Jana Amrichová and Jan Vondrácek and Stanislav Kozubek and Eva Bártová},

doi = {10.1002/jcb.22466},

issn = {1097-4644 0730-2312},

year  = {2010},

date = {2010-04-01},

journal = {Journal of cellular biochemistry},

volume = {109},

number = {5},

pages = {915–926},

abstract = {Telomeres are specialized chromatin structures that are situated at the end of linear chromosomes and play an important role in cell senescence and immortalization. Here, we investigated whether changes in histone signature influence the nuclear arrangement and positioning of telomeres. Analysis of mouse embryonic fibroblasts revealed that telomeres were organized into specific clusters that partially associated with centromeric clusters. This nuclear arrangement was influenced by deficiency of the histone methyltransferase SUV39h, LMNA deficiency, and the histone deacetylase inhibitor Trichostatin A (TSA). Similarly, nuclear radial distributions of telomeric clusters were preferentially influenced by TSA, which caused relocation of telomeres closer to the nuclear center. Telomeres also co-localized with promyelocytic leukemia bodies (PML). This association was increased by SUV39h deficiency and decreased by LMNA deficiency. These differences could be explained by differing levels of the telomerase subunit, TERT, in SUV39h- and LMNA-deficient fibroblasts. Taken together, our data show that SUV39h and A-type lamins likely play a key role in telomere maintenance and telomere nuclear architecture.},

note = {Place: United States},

keywords = {*Gene Rearrangement, Animals, DNA-Binding Proteins/metabolism, Epigenesis, Fibroblasts/metabolism, Flow Cytometry, Genetic, Humans, Intranuclear Inclusion Bodies/metabolism, Lamin Type A/*metabolism, Methyltransferases/*metabolism, Mice, Protein Transport, rap1 GTP-Binding Proteins/metabolism, Repressor Proteins/*metabolism, Shelterin Complex, Telomerase/metabolism, Telomere-Binding Proteins, Telomere/genetics/*metabolism, Telomeric Repeat Binding Protein 1/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}

}

@article{vanhara_growthdifferentiation_2009,

title = {Growth/differentiation factor-15 inhibits differentiation into osteoclasts–a novel factor involved in control of osteoclast differentiation.},

author = {Petr Vanhara and Eva Lincová and Alois Kozubík and Pierre Jurdic and Karel Soucek and Jan Smarda},

doi = {10.1016/j.diff.2009.07.008},

issn = {1432-0436 0301-4681},

year  = {2009},

date = {2009-11-01},

journal = {Differentiation; research in biological diversity},

volume = {78},

number = {4},

pages = {213–222},

abstract = {Survival and capability of cancer cells to form metastases fundamentally depend on interactions with their microenvironment. Secondary tumors originating from prostate carcinomas affect remodeling of bone tissue and can induce both osteolytic and osteocondensing lesions. However, particular molecular mechanisms responsible for selective homing and activity of cancer cells in bone microenvironment have not been clarified yet. Growth/differentiation factor-15 (GDF-15), a distant member of the TGF-beta protein family, has recently been associated with many human cancers, including prostate. We show that both pure GDF-15 and the GDF-15-containing growth medium of 1,25(OH)(2)-vitamin D(3)-treated prostate adenocarcinoma LNCaP cells suppress formation of mature osteoclasts differentiated from RAW264.7 macrophages and bone-marrow precursors by M-CSF/RANKL in a dose-dependent manner. GDF-15 inhibits expression of c-Fos and activity of NFkappaB by delayed degradation of IkappaB. Moreover, GDF-15 inhibits expression of carbonic anhydrase II and cathepsin K, key osteoclast enzymes, and induces changes in SMAD and p38 signaling. The lack of functional osteoclasts can contribute to accumulation of bone matrix by reduction of bone resorption. These results unveil new role of GDF-15 in modulation of osteoclast differentiation and possibly in therapy of bone metastases.},

note = {Place: England},

keywords = {Acid Phosphatase/metabolism, Animals, Calcitriol/pharmacology, Carbonic Anhydrase II/antagonists & inhibitors, Cathepsin K/antagonists & inhibitors/genetics, Cell Differentiation/*drug effects, Cell Line, Conditioned/pharmacology, Culture Media, Dose-Response Relationship, Drug, Femur/cytology, Growth Differentiation Factor 15/*pharmacology, Humans, Inbred Strains, Isoenzymes/metabolism, Macrophage Colony-Stimulating Factor/pharmacology, Macrophages/cytology, Male, Mice, NF-kappa B/antagonists & inhibitors, Osteoclasts/*drug effects/metabolism, Prostatic Neoplasms/metabolism, Proto-Oncogene Proteins c-fos/antagonists & inhibitors, RANK Ligand/pharmacology, Tartrate-Resistant Acid Phosphatase, Time Factors, Tumor},

pubstate = {published},

tppubtype = {article}

}

@article{takacova_role_2009,

title = {Role of aryl hydrocarbon receptor in modulation of the expression of the hypoxia marker carbonic anhydrase IX.},

author = {Martina Takacova and Tereza Holotnakova and Jan Vondracek and Miroslav Machala and Katerina Pencikova and Katarina Gradin and Lorenz Poellinger and Jaromir Pastorek and Silvia Pastorekova and Juraj Kopacek},

doi = {10.1042/BJ20080952},

issn = {1470-8728 0264-6021},

year  = {2009},

date = {2009-04-01},

journal = {The Biochemical journal},

volume = {419},

number = {2},

pages = {419–425},

abstract = {Tumour-associated expression of CA IX (carbonic anhydrase IX) is to a major extent regulated by HIF-1 (hypoxia-inducible factor-1) which is important for transcriptional activation and consists of the oxygen-regulated subunit HIF-1alpha and the partner factor ARNT [AhR (aryl hydrocarbon receptor) nuclear translocator]. We have previously observed that HIF-1alpha competes with the AhR for interaction with ARNT under conditions when both conditionally regulated factors are activated. We have therefore investigated whether TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin)-induced activation of the AhR pathway might interfere with CA IX expression. The results from the present study suggest that TCDD treatment reduces hypoxic induction of both CA IX mRNA and protein expression. Moreover, the transcriptional activity of the CA9 promoter was significantly reduced by expression of CAAhR (constitutively active AhR), which activates transcription in a ligand-independent manner. Finally, we found that ARNT is critical for both hypoxic induction and the TCDD-mediated inhibition of CA9 expression.},

note = {Place: England},

keywords = {alpha Subunit, Animals, Antigens, Aryl Hydrocarbon/genetics/metabolism/*physiology, Binding Sites, Blotting, Carbonic Anhydrase IX, Carbonic Anhydrases/genetics/*metabolism, Cell Hypoxia/genetics/*physiology, Cell Line, Chromatin Immunoprecipitation, Genetic/genetics, Humans, Hypoxia-Inducible Factor 1, Mice, Neoplasm/genetics, Polychlorinated Dibenzodioxins/pharmacology, Polymerase Chain Reaction, Promoter Regions, Protein Binding/drug effects, Receptors, Reverse Transcriptase Polymerase Chain Reaction, Signal Transduction/drug effects, Tumor, Western},

pubstate = {published},

tppubtype = {article}

}

@article{vistejnova_comparison_2009,

title = {The comparison of impedance-based method of cell proliferation monitoring with commonly used metabolic-based techniques.},

author = {Lucie Vistejnova and Jana Dvorakova and Martina Hasova and Tomas Muthny and Vladimir Velebny and Karel Soucek and Lukas Kubala},

issn = {0172-780X},

year  = {2009},

date = {2009-01-01},

journal = {Neuro endocrinology letters},

volume = {30 Suppl 1},

pages = {121–127},

abstract = {OBJECTIVES: Determination of cell numbers is a crucial step in studies focused on cytokinetics and cell toxicity. The impedance-based analysis employing electronic sensor array system xCELLigence System allowing label-free dynamic monitoring of relative viable adherent cell amounts was compared with the most utilized methods for relative quantification of viable cell numbers based on a determination of cellular metabolism. DESIGN: Colorimetric assay based on reduction of tetrazolium salt (MTT) by mitochondrial enzymes and chemiluminiscent assay based on intracellular adenosine triphosphate (ATP) determination were compared with the impedance-based system. Cell morphology was compared by microscopic evaluation. Normal human epidermal keratinocytes (NHEK) and normal human dermal fibroblasts (NHDF), together with 3T3 mouse fibroblast and HaCaT keratinocyte cell lines were employed. RESULTS: The progress of cell growth curves obtained by different methods during 72 hours reflected cell type and cell seeding densities. The impedance-based method was found to be applicable for the determination of the cell proliferation of 3T3 fibroblasts, HaCaT and NHDF, since the comparison of this method with ATP and MTT determinations showed a comparable results. In contrast, the proliferation of NHEK measured by the impedance-based method did not correlate with other methodological approaches. This could be accounted to the specific morphological appearance of these cells. CONCLUSION: The study shows the impedance-based detection of viable adherent cells is a valuable approach for cytokinetics and pharmacological studies. However, the specific morphological characteristics of cell lines have to be considered employing this method for determination of cell proliferation without using other reference methods.},

note = {Place: Sweden},

keywords = {*Cell Proliferation, *Electric Impedance, 3T3 Cells, Adenosine Triphosphate/*metabolism, Animals, Cell Count/*methods, Cell Line, Cells, Colorimetry, Cultured, Dermis/cytology, Fibroblasts/cytology, Humans, Keratinocytes/cytology, Luminescent Measurements, Mice, Mitochondria/enzymology/metabolism, Oxidation-Reduction, Tetrazolium Salts/*metabolism, Time Factors},

pubstate = {published},

tppubtype = {article}

}

@article{hoferova_vitro_2004,

title = {In vitro proliferation of fibrosarcoma cells depends on intact functions of lipoxygenases and cytochrome P-450-monooxygenase.},

author = {Zuzana Hoferová and Karel Soucek and Jirina Hofmanová and Michael Hofer and Katerina Chramostová and Peter Fedorocko and Alois Kozubik},

doi = {10.1081/cnv-120030212},

issn = {0735-7907},

year  = {2004},

date = {2004-01-01},

journal = {Cancer investigation},

volume = {22},

number = {2},

pages = {234–247},

abstract = {Proliferation of mouse fibrosarcoma cells G:5:113 was studied in vitro after affecting particular pathways of arachidonic acid metabolism by selected inhibitors. After 48 hours of cultivation with nonspecific lipoxygenase inhibitors, nordihydroguaiaretic acid (NDGA) and esculetin; a specific 12-lipoxygenase inhibitor, baicalein; and inhibitor of five-lipoxygenase activating protein, MK-886, markedly suppressed the number of cells and induced significant changes in cell cycle distribution in a dose-dependent manner. While proadifen, an inhibitor of cytochrome P-450-monooxygenase, applied in low concentrations, increased the cell number, at higher concentrations, it inhibited cell proliferation and significantly changed the cell cycle. Cyclooxygenase inhibitors, ibuprofen, flurbiprofen, and diclofenac suppressed cell numbers only moderately without any changes in the cell cycle. The occurrence of apoptosis was not significant for any of the selected drugs in comparison with untreated control cells. Moreover, not even one of the drugs caused the specific cleavage of poly (ADP-ribose) polymerase to the 89-kDa fragment, however, a decrease in total amount of this protein was observed after treatment with NDGA and esculetin. We conclude that the proliferation ability of fibrosarcoma cells G:5:113 in vitro depends on intact functions of 5-lipoxygenase, 12-lipoxygenase, and cytochrome P-450-monooxygenases, and that the effects of inhibitors do not include regulation of apoptosis.},

note = {Place: England},

keywords = {Animals, Apoptosis, Arachidonic Acid/antagonists & inhibitors/metabolism/*pharmacology, Cell Cycle/*physiology, Cultured, Cytochrome P-450 Enzyme System/*pharmacology, Enzyme Inhibitors/pharmacology, Fibrosarcoma/*pathology/veterinary, Lipoxygenase/*pharmacology, Mice, Tumor Cells},

pubstate = {published},

tppubtype = {article}

}

@article{hoferova_tumor-host_2003,

title = {Tumor-host interactions accompanying the growth of the G:5:113 fibrosarcoma in the mouse: possibilities for a new therapeutic approach?},

author = {Zuzana Hoferová and Antonín Vacek and Michal Hofer and Nadezda O. Macková and Karel Soucek and Alena Egyed and Peter Fedorocko},

doi = {10.1081/cnv-120016419},

issn = {0735-7907},

year  = {2003},

date = {2003-04-01},

journal = {Cancer investigation},

volume = {21},

number = {2},

pages = {227–236},

abstract = {The experiments were aimed at describing in detail some interactions between a solid tumor growing from subcutaneously transplanted G:5:113 fibrosarcoma cells in vivo and its mouse host. The tumor was found to elevate significantly the number of granulocytes in the peripheral blood of the host after having achieved the volume of about 1 cm3 (day 40 after transplantation). Blood plasma from fibrosarcoma-bearing mice stimulated proliferation of progenitor cells for granulocytes and macrophages (GM-CFC) in vitro and suppressed growth of G:5:113 cell population in culture. Interestingly, both effects were observable as early as week 1 when the tumor was still macroscopically invisible and unpalpable. Conditioned medium from cultures of G:5:113 fibrosarcoma cells stimulated proliferation of GM-CFC in vitro. These findings might represent a starting point for studies aimed at designing new therapeutic approaches for the treatment of fibrosarcoma.},

note = {Place: England},

keywords = {Animals, Cell Count, Cell Cycle/*physiology, Cell Division, Cell Survival/*physiology, Conditioned, Culture Media, Cultured, Fibrosarcoma/blood/*pathology, Granulocytes/pathology, Inbred C3H, Leukocyte Count, Mice, Tumor Cells},

pubstate = {published},

tppubtype = {article}

}

@article{hoferova_effect_2002,

title = {The effect of nonsteroidal antiinflammatory drugs ibuprofen, flurbiprofen, and diclofenac on in vitro and in vivo growth of mouse fibrosarcoma.},

author = {Zuzana Hoferová and Peter Fedorocko and Jirina Hofmanová and Michal Hofer and Vladimír Znojil and Katerina Minksová and Karel Soucek and Alena Egyed and Alois Kozubík},

doi = {10.1081/cnv-120002149},

issn = {0735-7907},

year  = {2002},

date = {2002-01-01},

journal = {Cancer investigation},

volume = {20},

number = {4},

pages = {490–498},

abstract = {For suppression of primary G:5:113 fibrosarcoma growth, three structurally different cyclooxygenase (COX) inhibitors (ibuprofen, flurbiprofen, and diclofenac) were administered intraperitoneally (i.p.) in two regimens starting on day 5 after tumor-cell inoculation. Repeated application of 0.15 mg/mouse/day during 14 consecutive days significantly suppressed the tumor growth and increased the percentage of surviving mice. Similar tendency, however without significant differences, was observed when animals were given 0.5 mg/day for five consecutive days. These results suggest that a time schedule of drug application is important for the therapeutic effect. Suppressive effect of diclofenac and flurbiprofen on tumor growth was also observed under in vitro conditions. We conclude that suppressive effect of these drugs on tumor growth in vivo comprises both direct effects of COX inhibitors on fibrosarcoma cells and indirect effects that are presumably mediated by extratumoral sources. Our findings encourage the use of COX inhibitors in the therapy of fibrosarcoma.},

note = {Place: England},

keywords = {Animals, Anti-Inflammatory Agents, Cell Cycle/drug effects, Cell Division/drug effects, Cultured/*drug effects, Diclofenac/*therapeutic use, Experimental, Fibrosarcoma/*drug therapy/pathology, Flurbiprofen/*therapeutic use, Ibuprofen/*therapeutic use, In Vitro Techniques, Inbred C3H, Male, Mice, Neoplasms, Non-Steroidal/*therapeutic use, Survival Rate, Tumor Cells},

pubstate = {published},

tppubtype = {article}

}