Latest Publications

@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_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{remsik_plasticity_2018,

title = {Plasticity and intratumoural heterogeneity of cell surface antigen expression in breast cancer.},

author = {Ján Remšík and Radek Fedr and Jiří Navrátil and Lucia Binó and Eva Slabáková and Pavel Fabian and Marek Svoboda and Karel Souček},

doi = {10.1038/bjc.2017.497},

issn = {1532-1827 0007-0920},

year  = {2018},

date = {2018-03-01},

journal = {British journal of cancer},

volume = {118},

number = {6},

pages = {813–819},

abstract = {Background:The intratumoural heterogeneity, often driven by epithelial-to-mesenchymal transition (EMT), significantly contributes to chemoresistance and disease progression in adenocarcinomas. Methods:We introduced a high-throughput screening platform to identify surface antigens that associate with epithelial–mesenchymal plasticity in well-defined pairs of epithelial cell lines and their mesenchymal counterparts. Using multicolour flow cytometry, we then analysed the expression of 10 most robustly changed antigens and identified a 10-molecule surface signature, in pan-cytokeratin-positive/EpCAM-positive and -negative fractions of dissociated breast tumours. Results:We found that surface CD9, CD29, CD49c, and integrin ß5 are lost in breast cancer cells that underwent EMT in vivo. The tetraspanin family member CD9 was concordantly downregulated both in vitro and in vivo and associated with epithelial phenotype and favourable prognosis. Conclusions:We propose that overall landscape of 10-molecule surface signature expression reflects the epithelial–mesenchymal plasticity in breast cancer.},

note = {Place: England},

keywords = {Antigens, Biomarkers, Breast Neoplasms/*genetics/*immunology/pathology, Cell Line, Cell Plasticity/immunology, Cellular Reprogramming/physiology, Epithelial-Mesenchymal Transition/immunology, Female, Flow Cytometry, Genetic, High-Throughput Screening Assays, Humans, Neoplasm Metastasis, Neoplasm/*biosynthesis/immunology, Surface/*biosynthesis/immunology, Tetraspanin 29/biosynthesis/immunology, Transcription, Tumor},

pubstate = {published},

tppubtype = {article}

}

@article{fedr_automatic_2013,

title = {Automatic cell cloning assay for determining the clonogenic capacity of cancer and cancer stem-like cells.},

author = {Radek Fedr and Zuzana Pernicová and Eva Slabáková and Nicol Straková and Jan Bouchal and Michal Grepl and Alois Kozubík and Karel Souček},

doi = {10.1002/cyto.a.22273},

issn = {1552-4930 1552-4922},

year  = {2013},

date = {2013-05-01},

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

volume = {83},

number = {5},

pages = {472–482},

abstract = {The clonogenic assay is a well-established in vitro method for testing the survival and proliferative capability of cells. It can be used to determine the cytotoxic effects of various treatments including chemotherapeutics and ionizing radiation. However, this approach can also characterize cells with different phenotypes and biological properties, such as stem cells or cancer stem cells. In this study, we implemented a faster and more precise method for assessing the cloning efficiency of cancer stem-like cells that were characterized and separated using a high-speed cell sorter. Cell plating onto a microplate using an automatic cell deposition unit was performed in a single-cell or dilution rank mode by the fluorescence-activated cell sorting method. We tested the new automatic cell-cloning assay (ACCA) on selected cancer cell lines and compared it with the manual approach. The obtained results were also compared with the results of the limiting dilution assay for different cell lines. We applied the ACCA to analyze the cloning capacity of different subpopulations of prostate and colon cancer cells based on the expression of the characteristic markers of stem (CD44 and CD133) and cancer stem cells (TROP-2, CD49f, and CD44). Our results revealed that the novel ACCA is a straightforward approach for determining the clonogenic capacity of cancer stem-like cells identified in both cell lines and patient samples.},

note = {Place: United States},

keywords = {*Cell Proliferation, AC133 Antigen, Antigens, Biomarkers, CD/metabolism, Cell Adhesion Molecules/metabolism, Cell Line, Cell Survival, Colonic Neoplasms/metabolism/*pathology, Flow Cytometry/*methods, Glycoproteins/metabolism, Humans, Hyaluronan Receptors/metabolism, In Vitro Techniques, Integrin alpha6/metabolism, Male, Neoplasm/metabolism, Neoplastic Stem Cells/metabolism/*pathology, Peptides/metabolism, Prostatic Neoplasms/metabolism/*pathology, Tumor, Tumor Stem Cell Assay/*methods, Tumor/metabolism},

pubstate = {published},

tppubtype = {article}

}

@article{benes_inhibition_2011,

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

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

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

issn = {1872-7980 0304-3835},

year  = {2011},

date = {2011-04-01},

journal = {Cancer letters},

volume = {303},

number = {1},

pages = {29–38},

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

note = {Place: Ireland},

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

pubstate = {published},

tppubtype = {article}

}

@article{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}

}