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
• Chromaffin Cells * metabolism   MeSH
• Catecholamines metabolism   MeSH
• Mice MeSH
• Adrenal Glands metabolism   MeSH
• Neuroblastoma * metabolism   MeSH
• Serotonin metabolism   MeSH
• Pregnancy MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Pregnancy MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Catecholamines 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.

• Keywords
• all-trans retinoic acid, calcidiol, calcitriol, osteosarcoma, retinoic acid receptor α, vitamin D receptor,
• MeSH
• Retinoic Acid Receptor alpha metabolism   MeSH
• Calcifediol administration & dosage   MeSH
• Calcitriol administration & dosage   MeSH
• Humans MeSH
• Cell Line, Tumor MeSH
• Osteosarcoma drug therapy   metabolism   MeSH
• Antineoplastic Agents administration & dosage   MeSH
• Antineoplastic Combined Chemotherapy Protocols MeSH
• Receptors, Calcitriol metabolism   MeSH
• Drug Screening Assays, Antitumor MeSH
• Tretinoin administration & dosage   MeSH
• Vitamins administration & dosage   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Retinoic Acid Receptor alpha MeSH
• Calcifediol MeSH
• Calcitriol MeSH
• Antineoplastic Agents MeSH
• Receptors, Calcitriol MeSH
• Tretinoin MeSH
• Vitamins 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
• Bexarotene pharmacology   MeSH
• Biomarkers, Pharmacological metabolism   MeSH
• Drug Resistance, Neoplasm drug effects   genetics   MeSH
• DEAD-box RNA Helicases genetics   metabolism   MeSH
• Child MeSH
• Fenretinide pharmacology   MeSH
• Tissue Fixation MeSH
• Homeodomain Proteins genetics   metabolism   MeSH
• Isotretinoin pharmacology   MeSH
• Infant MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Cell Line, Tumor MeSH
• Nervous System Neoplasms genetics   metabolism   pathology   surgery   MeSH
• Neuroblastoma genetics   metabolism   pathology   surgery   MeSH
• Infant, Newborn MeSH
• Pre-B-Cell Leukemia Transcription Factor 1 genetics   metabolism   MeSH
• Child, Preschool MeSH
• Cell Proliferation drug effects   MeSH
• HMGA1a Protein genetics   metabolism   MeSH
• HMGA2 Protein genetics   metabolism   MeSH
• Antineoplastic Agents pharmacology   MeSH
• Tretinoin analogs & derivatives   pharmacology   MeSH
• Paraffin Embedding MeSH
• Check Tag
• Child MeSH
• Infant MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Male MeSH
• Infant, Newborn MeSH
• Child, Preschool MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• 9,13-retinoic acid MeSH Browser
• Bexarotene MeSH
• Biomarkers, Pharmacological MeSH
• DDX39A protein, human MeSH Browser
• DEAD-box RNA Helicases MeSH
• Fenretinide MeSH
• HMGA2 protein, human MeSH Browser
• Homeodomain Proteins MeSH
• Hoxc9 protein, human MeSH Browser
• Isotretinoin MeSH
• PBX1 protein, human MeSH Browser
• Pre-B-Cell Leukemia Transcription Factor 1 MeSH
• HMGA1a Protein MeSH
• HMGA2 Protein MeSH
• Antineoplastic Agents 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.

• Keywords
• FR180204, U0126, erlotinib, infantile myofibromatosis, platelet-derived growth factor receptor, protein kinase inhibitors, receptor tyrosine kinases, sunitinib, targeted therapy,
• MeSH
• Butadienes administration & dosage   therapeutic use   MeSH
• Child MeSH
• Erlotinib Hydrochloride administration & dosage   therapeutic use   MeSH
• Phosphorylation drug effects   MeSH
• Protein Kinase Inhibitors administration & dosage   therapeutic use   MeSH
• Infant MeSH
• Humans MeSH
• Mutation * MeSH
• Myofibromatosis congenital   drug therapy   genetics   MeSH
• Cell Line, Tumor MeSH
• Nitriles administration & dosage   therapeutic use   MeSH
• Cell Proliferation drug effects   MeSH
• Pyrazoles administration & dosage   therapeutic use   MeSH
• Pyridazines administration & dosage   therapeutic use   MeSH
• Receptor, Platelet-Derived Growth Factor beta * genetics   metabolism   MeSH
• Sunitinib administration & dosage   therapeutic use   MeSH
• Check Tag
• Child MeSH
• Infant MeSH
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Butadienes MeSH
• Erlotinib Hydrochloride MeSH
• FR 180204 MeSH Browser
• Protein Kinase Inhibitors MeSH
• Nitriles MeSH
• PDGFRB protein, human MeSH Browser
• Pyrazoles MeSH
• Pyridazines MeSH
• Receptor, Platelet-Derived Growth Factor beta * MeSH
• Sunitinib MeSH
• U 0126 MeSH Browser

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
• Child MeSH
• Adult MeSH
• Fluorescent Antibody Technique MeSH
• Immunoblotting MeSH
• Immunohistochemistry MeSH
• Infant MeSH
• Humans MeSH
• Medulloblastoma pathology   MeSH
• Adolescent MeSH
• Young Adult MeSH
• Cerebellum pathology   MeSH
• Biomarkers, Tumor analysis   MeSH
• Tumor Cells, Cultured MeSH
• Cerebellar Neoplasms pathology   MeSH
• Child, Preschool MeSH
• Check Tag
• Child MeSH
• Adult MeSH
• Infant MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Male MeSH
• Child, Preschool MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Biomarkers, Tumor 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
• Adenocarcinoma diagnosis   metabolism   MeSH
• Epithelial Cell Adhesion Molecule metabolism   MeSH
• AC133 Antigen metabolism   MeSH
• CD24 Antigen metabolism   MeSH
• Hyaluronan Receptors metabolism   MeSH
• Stem Cells metabolism   MeSH
• Cells, Cultured MeSH
• Real-Time Polymerase Chain Reaction MeSH
• Middle Aged MeSH
• Humans MeSH
• Biomarkers, Tumor MeSH
• Pancreatic Neoplasms diagnosis   metabolism   MeSH
• Nestin metabolism   MeSH
• Prognosis MeSH
• Flow Cytometry MeSH
• Gene Expression Regulation, Neoplastic MeSH
• Aged MeSH
• Transcriptome MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Epithelial Cell Adhesion Molecule MeSH
• AC133 Antigen MeSH
• CD24 Antigen MeSH
• Hyaluronan Receptors MeSH
• EPCAM protein, human MeSH Browser
• Biomarkers, Tumor MeSH
• NES protein, human MeSH Browser
• Nestin MeSH
• PROM1 protein, human MeSH Browser

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.

• Keywords
• Cancer stem cells, Markers, Pediatric sarcomas, Sox2, Tumorigenicity,
• MeSH
• ATP Binding Cassette Transporter, Subfamily G, Member 2 metabolism   MeSH
• AC133 Antigen metabolism   MeSH
• Child MeSH
• Adult MeSH
• Humans MeSH
• Adolescent MeSH
• Mice, Inbred NOD MeSH
• Mice, SCID MeSH
• Mice MeSH
• Cell Transformation, Neoplastic metabolism   pathology   MeSH
• Biomarkers, Tumor metabolism   MeSH
• Cell Line, Tumor MeSH
• Neoplastic Stem Cells metabolism   pathology   MeSH
• Neoplasm Proteins metabolism   MeSH
• Nestin metabolism   MeSH
• Osteosarcoma metabolism   pathology   MeSH
• Child, Preschool MeSH
• Rhabdomyosarcoma metabolism   pathology   MeSH
• Sarcoma metabolism   pathology   MeSH
• SOXB1 Transcription Factors metabolism   MeSH
• Animals MeSH
• Check Tag
• Child MeSH
• Adult MeSH
• Humans MeSH
• Adolescent MeSH
• Male MeSH
• Mice MeSH
• Child, Preschool MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH
• Names of Substances
• ATP Binding Cassette Transporter, Subfamily G, Member 2 MeSH
• ABCG2 protein, human MeSH Browser
• AC133 Antigen MeSH
• Biomarkers, Tumor MeSH
• Neoplasm Proteins MeSH
• Nestin MeSH
• PROM1 protein, human MeSH Browser
• SOX2 protein, human MeSH Browser
• SOXB1 Transcription Factors 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.

• Keywords
• Cancer stem cells, cytoskeleton, intermediate filaments, nestin, tumor markers,
• MeSH
• Humans MeSH
• Biomarkers, Tumor metabolism   MeSH
• Neoplastic Stem Cells metabolism   MeSH
• Neoplasms metabolism   pathology   MeSH
• Nestin metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Names of Substances
• Biomarkers, Tumor MeSH
• NES protein, human MeSH Browser
• 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
• AC133 Antigen MeSH
• Cell Nucleus metabolism   MeSH
• Antigens, CD immunology   metabolism   MeSH
• Fluorescent Antibody Technique, Indirect MeSH
• Glycoproteins immunology   metabolism   MeSH
• Humans MeSH
• Antibodies, Monoclonal immunology   MeSH
• Biomarkers, Tumor metabolism   MeSH
• Cell Line, Tumor MeSH
• Peptides immunology   metabolism   MeSH
• Rhabdomyosarcoma metabolism   MeSH
• Microscopy, Electron, Transmission MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• AC133 Antigen MeSH
• Antigens, CD MeSH
• Glycoproteins MeSH
• Antibodies, Monoclonal MeSH
• Biomarkers, Tumor MeSH
• Peptides MeSH
• PROM1 protein, human MeSH Browser
• Prom1 protein, mouse MeSH Browser

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
• Cell Nucleus chemistry   metabolism   ultrastructure   MeSH
• Child MeSH
• Fluorescent Antibody Technique MeSH
• Glioblastoma metabolism   ultrastructure   MeSH
• Immunohistochemistry MeSH
• Infant MeSH
• Middle Aged MeSH
• Humans MeSH
• Medulloblastoma metabolism   ultrastructure   MeSH
• Cell Line, Tumor MeSH
• Nestin MeSH
• Neuroblastoma metabolism   ultrastructure   MeSH
• Child, Preschool MeSH
• Intermediate Filament Proteins analysis   metabolism   MeSH
• Nerve Tissue Proteins analysis   metabolism   MeSH
• Aged MeSH
• Microscopy, Electron, Transmission MeSH
• Blotting, Western MeSH
• Check Tag
• Child MeSH
• Infant MeSH
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Child, Preschool MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• NES protein, human MeSH Browser
• Nestin MeSH
• Intermediate Filament Proteins MeSH
• Nerve Tissue Proteins MeSH