Adrenal glands are the major organs releasing catecholamines and regulating our stress response. The mechanisms balancing generation of adrenergic chromaffin cells and protecting against neuroblastoma tumors are still enigmatic. Here we revealed that serotonin (5HT) controls the numbers of chromaffin cells by acting upon their immediate progenitor "bridge" cells via 5-hydroxytryptamine receptor 3A (HTR3A), and the aggressive HTR3Ahigh human neuroblastoma cell lines reduce proliferation in response to HTR3A-specific agonists. In embryos (in vivo), the physiological increase of 5HT caused a prolongation of the cell cycle in "bridge" progenitors leading to a smaller chromaffin population and changing the balance of hormones and behavioral patterns in adulthood. These behavioral effects and smaller adrenals were mirrored in the progeny of pregnant female mice subjected to experimental stress, suggesting a maternal-fetal link that controls developmental adaptations. Finally, these results corresponded to a size-distribution of adrenals found in wild rodents with different coping strategies.

• MeSH
• chromafinní buňky * metabolismus   MeSH
• katecholaminy metabolismus   MeSH
• myši MeSH
• nadledviny metabolismus   MeSH
• neuroblastom * metabolismus   MeSH
• serotonin metabolismus   MeSH
• těhotenství MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• katecholaminy MeSH
• serotonin MeSH

The main objective of this study was to analyze changes in the antiproliferative effect of vitamin D3, in the form of calcitriol and calcidiol, via its combined application with all-trans retinoic acid (ATRA) in osteosarcoma cell lines. The response to treatment with calcitriol and calcidiol alone was specific for each cell line. Nevertheless, we observed an enhanced effect of combined treatment with ATRA and calcitriol in the majority of the cell lines. Although the levels of respective nuclear receptors did not correlate with the sensitivity of cells to these drugs, vitamin D receptor (VDR) upregulation induced by ATRA was found in cell lines that were the most sensitive to the combined treatment. In addition, all these cell lines showed high endogenous levels of retinoic acid receptor α (RARα). Our study confirmed that the combination of calcitriol and ATRA can achieve enhanced antiproliferative effects in human osteosarcoma cell lines in vitro. Moreover, we provide the first evidence that ATRA is able to upregulate VDR expression in human osteosarcoma cells. According to our results, the endogenous levels of RARα and VDR could be used as a predictor of possible synergy between ATRA and calcitriol in osteosarcoma cells.

• Klíčová slova
• all-trans retinoic acid, calcidiol, calcitriol, osteosarcoma, retinoic acid receptor α, vitamin D receptor,
• MeSH
• alfa receptor kyseliny retinové metabolismus   MeSH
• kalcifediol aplikace a dávkování   MeSH
• kalcitriol aplikace a dávkování   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• osteosarkom farmakoterapie   metabolismus   MeSH
• protinádorové látky aplikace a dávkování   MeSH
• protokoly protinádorové kombinované chemoterapie MeSH
• receptory kalcitriolu metabolismus   MeSH
• screeningové testy protinádorových léčiv MeSH
• tretinoin aplikace a dávkování   MeSH
• vitaminy aplikace a dávkování   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• alfa receptor kyseliny retinové MeSH
• kalcifediol MeSH
• kalcitriol MeSH
• protinádorové látky MeSH
• receptory kalcitriolu MeSH
• tretinoin MeSH
• vitaminy MeSH

Although the administration of retinoids represents an important part of treatment for children suffering from high-risk neuroblastomas, approximately 50% of these patients do not respond to this therapy or develop resistance to retinoids during treatment. Our study focused on the comparative analysis of the expression of five genes and corresponding proteins (DDX39A, HMGA1, HMGA2, HOXC9 and PBX1) that have recently been discussed as possible predictive biomarkers of clinical response to retinoid differentiation therapy. Expression of these five candidate biomarkers was evaluated at both the mRNA and protein level in the same subset of 8 neuroblastoma cell lines after treatment with natural or synthetic retinoids. We found that the cell lines that were HMGA2-positive and/or HOXC9-negative have a reduced sensitivity to retinoids. Furthermore, the experiments revealed that the retinoid-sensitive cell lines showed a uniform pattern of change after treatment with both natural and sensitive retinoids: increased DDX39A and decreased PBX1 protein levels. Our results showed that in NBL cells, these putative protein biomarkers are associated with sensitivity or resistance to retinoids, and their endogenous or induced expression can distinguish between these two phenotypes.

• MeSH
• bexaroten farmakologie   MeSH
• biomarkery farmakologické metabolismus   MeSH
• chemorezistence účinky léků   genetika   MeSH
• DEAD-box RNA-helikasy genetika   metabolismus   MeSH
• dítě MeSH
• fenretinid farmakologie   MeSH
• fixace tkání MeSH
• homeodoménové proteiny genetika   metabolismus   MeSH
• isotretinoin farmakologie   MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• nádorové buněčné linie MeSH
• nádory nervového systému genetika   metabolismus   patologie   chirurgie   MeSH
• neuroblastom genetika   metabolismus   patologie   chirurgie   MeSH
• novorozenec MeSH
• pre-B-buněčný leukemický transkripční faktor 1 genetika   metabolismus   MeSH
• předškolní dítě MeSH
• proliferace buněk účinky léků   MeSH
• protein HMGA1A genetika   metabolismus   MeSH
• protein HMGA2 genetika   metabolismus   MeSH
• protinádorové látky farmakologie   MeSH
• tretinoin analogy a deriváty   farmakologie   MeSH
• zalévání tkání do parafínu MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 9,13-retinoic acid MeSH Prohlížeč
• bexaroten MeSH
• biomarkery farmakologické MeSH
• DDX39A protein, human MeSH Prohlížeč
• DEAD-box RNA-helikasy MeSH
• fenretinid MeSH
• HMGA2 protein, human MeSH Prohlížeč
• homeodoménové proteiny MeSH
• Hoxc9 protein, human MeSH Prohlížeč
• isotretinoin MeSH
• PBX1 protein, human MeSH Prohlížeč
• pre-B-buněčný leukemický transkripční faktor 1 MeSH
• protein HMGA1A MeSH
• protein HMGA2 MeSH
• protinádorové látky MeSH
• tretinoin MeSH

Infantile myofibromatosis represents one of the most common proliferative fibrous tumors of infancy and childhood. More effective treatment is needed for drug-resistant patients, and targeted therapy using specific protein kinase inhibitors could be a promising strategy. To date, several studies have confirmed a connection between the p.R561C mutation in gene encoding platelet-derived growth factor receptor beta (PDGFR-beta) and the development of infantile myofibromatosis. This study aimed to analyze the phosphorylation of important kinases in the NSTS-47 cell line derived from a tumor of a boy with infantile myofibromatosis who harbored the p.R561C mutation in PDGFR-beta. The second aim of this study was to investigate the effects of selected protein kinase inhibitors on cell signaling and the proliferative activity of NSTS-47 cells. We confirmed that this tumor cell line showed very high phosphorylation levels of PDGFR-beta, extracellular signal-regulated kinases (ERK) 1/2 and several other protein kinases. We also observed that PDGFR-beta phosphorylation in tumor cells is reduced by the receptor tyrosine kinase inhibitor sunitinib. In contrast, MAPK/ERK kinases (MEK) 1/2 and ERK1/2 kinases remained constitutively phosphorylated after treatment with sunitinib and other relevant protein kinase inhibitors. Our study showed that sunitinib is a very promising agent that affects the proliferation of tumor cells with a p.R561C mutation in PDGFR-beta.

• Klíčová slova
• FR180204, U0126, erlotinib, infantile myofibromatosis, platelet-derived growth factor receptor, protein kinase inhibitors, receptor tyrosine kinases, sunitinib, targeted therapy,
• MeSH
• butadieny aplikace a dávkování   terapeutické užití   MeSH
• dítě MeSH
• erlotinib aplikace a dávkování   terapeutické užití   MeSH
• fosforylace účinky léků   MeSH
• inhibitory proteinkinas aplikace a dávkování   terapeutické užití   MeSH
• kojenec MeSH
• lidé MeSH
• mutace * MeSH
• myofibromatóza vrozené   farmakoterapie   genetika   MeSH
• nádorové buněčné linie MeSH
• nitrily aplikace a dávkování   terapeutické užití   MeSH
• proliferace buněk účinky léků   MeSH
• pyrazoly aplikace a dávkování   terapeutické užití   MeSH
• pyridaziny aplikace a dávkování   terapeutické užití   MeSH
• růstový faktor odvozený z trombocytů - receptor beta * genetika   metabolismus   MeSH
• sunitinib aplikace a dávkování   terapeutické užití   MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• butadieny MeSH
• erlotinib MeSH
• FR 180204 MeSH Prohlížeč
• inhibitory proteinkinas MeSH
• nitrily MeSH
• PDGFRB protein, human MeSH Prohlížeč
• pyrazoly MeSH
• pyridaziny MeSH
• růstový faktor odvozený z trombocytů - receptor beta * MeSH
• sunitinib MeSH
• U 0126 MeSH Prohlížeč

Medulloblastoma comprises four main subgroups (WNT, SHH, Group 3 and Group 4) originally defined by transcriptional profiling. In primary medulloblastoma tissues, these groups are thought to be distinguishable using the immunohistochemical detection of β-catenin, filamin A, GAB1 and YAP1 protein markers. To investigate the utility of these markers for in vitro studies using medulloblastoma cell lines, immunoblotting and indirect immunofluorescence were employed for the detection of β-catenin, filamin A, GAB1 and YAP1 in both DAOY and D283 Med reference cell lines and the panel of six medulloblastoma cell lines derived in our laboratory from the primary tumor tissues of known molecular subgroups. Immunohistochemical detection of these markers was performed on formalin-fixed paraffin-embedded tissue of the matching primary tumors. The results revealed substantial divergences between the primary tumor tissues and matching cell lines in the immunoreactivity pattern of medulloblastoma-subgroup-specific protein markers. Regardless of the molecular subgroup of the primary tumor, all six patient-derived medulloblastoma cell lines exhibited a uniform phenotype: immunofluorescence showed the nuclear localization of YAP1, accompanied by strong cytoplasmic positivity for β-catenin and filamin A, as well as weak positivity for GAB1. The same immunoreactivity pattern was also found in both DAOY and D283 Med reference medulloblastoma cell lines. Therefore, we can conclude that various medulloblastoma cell lines tend to exhibit the same characteristics of protein marker expression under standard in vitro conditions. Such a finding emphasizes the importance of the analyses of primary tumors in clinically oriented medulloblastoma research and the urgent need to develop in vitro models of improved clinical relevance, such as 3D cultures and organotypic slice cultures.

• MeSH
• dítě MeSH
• dospělí MeSH
• fluorescenční protilátková technika MeSH
• imunoblotting MeSH
• imunohistochemie MeSH
• kojenec MeSH
• lidé MeSH
• meduloblastom patologie   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mozeček patologie   MeSH
• nádorové biomarkery analýza   MeSH
• nádorové buňky kultivované MeSH
• nádory mozečku patologie   MeSH
• předškolní dítě MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• nádorové biomarkery MeSH

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies. Its dismal prognosis is often attributed to the presence of cancer stem cells (CSCs) that have been identified in PDAC using various markers. However, the co-expression of all of these markers has not yet been evaluated. Furthermore, studies that compare the expression levels of CSC markers in PDAC tumor samples and in cell lines derived directly from those tumors are lacking. Here, we analyzed the expression of putative CSC markers-CD24, CD44, epithelial cell adhesion molecule (EpCAM), CD133, and nestin-by immunofluorescence, flow cytometry and quantitative PCR in 3 PDAC-derived cell lines and by immunohistochemistry in 3 corresponding tumor samples. We showed high expression of the examined CSC markers among all of the cell lines and tumor samples, with the exception of CD24 and CD44, which were enriched under in vitro conditions compared with tumor tissues. The proportions of cells positive for the remaining markers were comparable to those detected in the corresponding tumors. Co-expression analysis using flow cytometry revealed that CD24+/CD44+/EpCAM+/CD133+ cells represented a significant population of the cells (range, 43 to 72%) among the cell lines. The highest proportion of CD24+/CD44+/EpCAM+/CD133+ cells was detected in the cell line derived from the tumor of a patient with the shortest survival. Using gene expression profiling, we further identified the specific pro-tumorigenic expression profile of this cell line compared with the profiles of the other two cell lines. Together, CD24+/CD44+/EpCAM+/CD133+ cells are present in PDAC cell lines derived from primary tumors, and their increased proportion corresponds with a pro-tumorigenic gene expression profile.

• MeSH
• adenokarcinom diagnóza   metabolismus   MeSH
• adhezní molekula epiteliálních buněk metabolismus   MeSH
• antigen AC133 metabolismus   MeSH
• antigen CD24 metabolismus   MeSH
• antigeny CD44 metabolismus   MeSH
• kmenové buňky metabolismus   MeSH
• kultivované buňky MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové biomarkery MeSH
• nádory slinivky břišní diagnóza   metabolismus   MeSH
• nestin metabolismus   MeSH
• prognóza MeSH
• průtoková cytometrie MeSH
• regulace genové exprese u nádorů MeSH
• senioři MeSH
• transkriptom MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adhezní molekula epiteliálních buněk MeSH
• antigen AC133 MeSH
• antigen CD24 MeSH
• antigeny CD44 MeSH
• EPCAM protein, human MeSH Prohlížeč
• nádorové biomarkery MeSH
• NES protein, human MeSH Prohlížeč
• nestin MeSH
• PROM1 protein, human MeSH Prohlížeč

The three most frequent pediatric sarcomas, i.e., Ewing's sarcoma, osteosarcoma, and rhabdomyosarcoma, were examined in this study: three cell lines derived from three primary tumor samples were analyzed from each of these tumor types. Detailed comparative analysis of the expression of three putative cancer stem cell markers related to sarcomas-ABCG2, CD133, and nestin-was performed on both primary tumor tissues and corresponding cell lines. The obtained results showed that the frequency of ABCG2-positive and CD133-positive cells was predominantly increased in the respective cell lines but that the high levels of nestin expression were reduced in both osteosarcomas and rhabdomyosarcomas under in vitro conditions. These findings suggest the selection advantage of cells expressing ABCG2 or CD133, but the functional tests in NOD/SCID gamma mice did not confirm the tumorigenic potential of cells harboring this phenotype. Subsequent analysis of the expression of common stem cell markers revealed an evident relationship between the expression of the transcription factor Sox2 and the tumorigenicity of the cell lines in immunodeficient mice: the Sox2 levels were highest in the two cell lines that were demonstrated as tumorigenic. Furthermore, Sox2-positive cells were found in the respective primary tumors and all xenograft tumors showed apparent accumulation of these cells. All of these findings support our conclusion that regardless of the expression of ABCG2, CD133 and nestin, only cells displaying increased Sox2 expression are directly involved in tumor initiation and growth; therefore, these cells fit the definition of the cancer stem cell phenotype.

• Klíčová slova
• Cancer stem cells, Markers, Pediatric sarcomas, Sox2, Tumorigenicity,
• MeSH
• ABC transportér z rodiny G, člen 2 metabolismus   MeSH
• antigen AC133 metabolismus   MeSH
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• myši inbrední NOD MeSH
• myši SCID MeSH
• myši MeSH
• nádorová transformace buněk metabolismus   patologie   MeSH
• nádorové biomarkery metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádorové kmenové buňky metabolismus   patologie   MeSH
• nádorové proteiny metabolismus   MeSH
• nestin metabolismus   MeSH
• osteosarkom metabolismus   patologie   MeSH
• předškolní dítě MeSH
• rhabdomyosarkom metabolismus   patologie   MeSH
• sarkom metabolismus   patologie   MeSH
• transkripční faktory SOXB1 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• myši MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• ABC transportér z rodiny G, člen 2 MeSH
• ABCG2 protein, human MeSH Prohlížeč
• antigen AC133 MeSH
• nádorové biomarkery MeSH
• nádorové proteiny MeSH
• nestin MeSH
• PROM1 protein, human MeSH Prohlížeč
• SOX2 protein, human MeSH Prohlížeč
• transkripční faktory SOXB1 MeSH

The crucial role of cancer stem cells (CSCs) in the pathology of malignant diseases has been extensively studied during the last decade. Nestin, a class VI intermediate filament protein, was originally detected in neural stem cells during development. Its expression has also been reported in different tissues under various pathological conditions. Specifically, nestin has been shown to be expressed in transformed cells of various human malignancies, and a correlation between its expression and the clinical course of some diseases has been proved. Furthermore, the coexpression of nestin with other stem cell markers was described as a CSC phenotype that was subsequently verified using tumorigenicity assays. The primary aim of this review is to summarize the recent findings regarding nestin expression in CSCs, its possible role in CSC phenotypes, particularly with respect to capacity for self-renewal, and its utility as a putative marker of CSCs.

• Klíčová slova
• Cancer stem cells, cytoskeleton, intermediate filaments, nestin, tumor markers,
• MeSH
• lidé MeSH
• nádorové biomarkery metabolismus   MeSH
• nádorové kmenové buňky metabolismus   MeSH
• nádory metabolismus   patologie   MeSH
• nestin metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• nádorové biomarkery MeSH
• NES protein, human MeSH Prohlížeč
• nestin MeSH

CD133 (also known as prominin-1) is a cell surface glycoprotein that is widely used for the identification of stem cells. Furthermore, its glycosylated epitope, AC133, has recently been discussed as a marker of cancer stem cells in various human malignancies. During our recent experiments on rhabdomyosarcomas (RMS), we unexpectedly identified an atypical nuclear localization of CD133 in a relatively stable subset of cells in five RMS cell lines established in our laboratory. To the best of our knowledge, this atypical localization of CD133 has not yet been proven or analyzed in detail in cancer cells. In the present study, we verified the nuclear localization of CD133 in RMS cells using three independent anti-CD133 antibodies, including both rabbit polyclonal and mouse monoclonal antibodies. Indirect immunofluorescence and confocal microscopy followed by software cross-section analysis, transmission electron microscopy and cell fractionation with immunoblotting were also employed, and all the results undeniably confirmed the presence of CD133 in the nuclei of stable minor subpopulations of all five RMS cell lines. The proportion of cells showing an exclusive nuclear localization of CD133 ranged from 3.4 to 7.5%, with only minor differences observed among the individual anti-CD133 antibodies. Although the role of CD133 in the cell nucleus remains unclear, these results clearly indicate that this atypical nuclear localization of CD133 in a minor subpopulation of cancer cells is a common phenomenon in RMS cell lines.

• MeSH
• antigen AC133 MeSH
• buněčné jádro metabolismus   MeSH
• CD antigeny imunologie   metabolismus   MeSH
• fluorescenční protilátková technika nepřímá MeSH
• glykoproteiny imunologie   metabolismus   MeSH
• lidé MeSH
• monoklonální protilátky imunologie   MeSH
• nádorové biomarkery metabolismus   MeSH
• nádorové buněčné linie MeSH
• peptidy imunologie   metabolismus   MeSH
• rhabdomyosarkom metabolismus   MeSH
• transmisní elektronová mikroskopie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigen AC133 MeSH
• CD antigeny MeSH
• glykoproteiny MeSH
• monoklonální protilátky MeSH
• nádorové biomarkery MeSH
• peptidy MeSH
• PROM1 protein, human MeSH Prohlížeč
• Prom1 protein, mouse MeSH Prohlížeč

Nestin is a class VI intermediate filament protein expressed in the cytoplasm of stem and progenitor cells in the mammalian CNS during development. In adults, nestin is present only in a small subset of cells and tissues, including the subventricular zone of the adult mammalian brain, where neurogenesis occurs. Nestin expression has also been detected under such pathological conditions as ischemia, inflammation, and brain injury, as well as in various types of human solid tumors and their corresponding cell lines. Furthermore, nestin was recently found in the nuclei of glioblastoma, neuroblastoma, and angiosarcoma cells and it was proved to interact directly with the nuclear DNA in neuroblastoma cells. Here, we perform the first study of the intracellular distribution of nestin in cell lines derived from neurogenic tumors. Using immunodetection methods, we examined nestin expression in tumor-derived cell lines obtained from 11 patients with neuroblastoma, medulloblastoma, or glioblastoma multiforme. Besides its standard cytoplasmic localization, nestin was present in the nuclei of two neuroblastoma cell lines and one medulloblastoma cell line. Nestin was only present in the nuclei of cells with diffuse cytoplasmic staining for this protein, and the proportion of cells positive for nestin in nuclei, as well as the intensity of staining, varied. The presence of nestin in the nuclei was confirmed by both transmission electron microscopy and Western blotting. Our results indicate that the presence of nestin in the nuclei of tumor cells is not very rare, especially under in vitro conditions.

• MeSH
• buněčné jádro chemie   metabolismus   ultrastruktura   MeSH
• dítě MeSH
• fluorescenční protilátková technika MeSH
• glioblastom metabolismus   ultrastruktura   MeSH
• imunohistochemie MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• meduloblastom metabolismus   ultrastruktura   MeSH
• nádorové buněčné linie MeSH
• nestin MeSH
• neuroblastom metabolismus   ultrastruktura   MeSH
• předškolní dítě MeSH
• proteiny intermediálních filament analýza   metabolismus   MeSH
• proteiny nervové tkáně analýza   metabolismus   MeSH
• senioři MeSH
• transmisní elektronová mikroskopie MeSH
• western blotting MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• NES protein, human MeSH Prohlížeč
• nestin MeSH
• proteiny intermediálních filament MeSH
• proteiny nervové tkáně MeSH