RESUMEN
BACKGROUND: Maffucci syndrome is a rare congenital nonhereditary disease characterized by multiple hemangiomas and enchondromas, which may progress into malignancy. The causal therapy does not exist, and therapy is aimed at complications. The determination of appropriate therapy is complicated, and a multidisciplinary approach is often essential. CASE: Authors are presenting the case of a 20-yearâ-old patient with Maffucci syndrome. During her life, multiple enchondromas and progressing hemangiomas have been revealed and they have caused many complications, such as limited movement, growth failure, pain, fluidothorax and ascites. A profile of phosphorylation of selected tyrosine kinases and MAP kinases from progressing hemangioma was performed and with consideration of the result, it led to change of treatment strategy with encouraging clinical response lasting for six months.
Asunto(s)
Encondromatosis/terapia , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Proteínas Tirosina Quinasas/metabolismo , Adulto , Activación Enzimática , Femenino , Humanos , FosforilaciónRESUMEN
Cancer stem cells (CSCs) are considered to be a population of tumor cells, which are responsible for tumor initiation and progression. They are also involved in metastasizing and may be a possible cause of multidrug resistance and tumor recurrence. CSCs possess the ability to self ârenew and show a tumorigenic potential. Functional assays, which enable the detection of these properties, represent the main tool for identification of CSCs. This article summarizes both in vitro and in vivo methods used to identify the CSCs with emphasis on recently employed techniques of CSCs detection. In vivo tumorigenicity assay, sphere formation assay and colony âforming unit assay belong to the most commonly used functional assays. Further, label âretention assay and aldehyde dehydrogenase activity assay are described in this article.
Asunto(s)
Células Madre Neoplásicas/metabolismo , Animales , Pruebas de Carcinogenicidad , Técnicas Citológicas , Humanos , Ratones , Ensayo de Tumor de Célula MadreRESUMEN
Tumorigenesis is always accompanied by alterations of the microenvironment in the respective tissue. The tumor microenvironment represents a heterogeneous complex, in which cell proliferation, differentiation, necrosis or apoptosis are regulated by various extracellular stimuli, and it can also lead to development of an aggressive phenotype of tumor cells. Influence of tumor microenvironment is also often connected with resistance to frequently used therapeutic procedures. Specifics of the tumor microenvironment are closely associated with the structural and functional abnormalities of tumor microvessels and altered cellular metabolism. Moreover, changes such as increase in glycolysis, elevated glucose uptake, production of lactate and CO2, and presence of hypoxic regions and regions with acidic pH are typical features of tumor tissues. At present, there is a lot of methods for in vitro simulation and investigation of some of these specific conditions, and a number of new methods are being developed. A detailed understanding of the specifics of the tumor microenvironment should increasingly improve the development of new treatment possibilities of human cancers.
Asunto(s)
Técnicas In Vitro , Microambiente Tumoral , HumanosRESUMEN
The identification of cancer stem cell markers represents one of the very relevant research topics because cancer stem cells play important roles in tumour initiation and progression, as well as during metastasis formation and in relapse of the disease. This article summarises recent knowledge on well-known and putative cancer stem cell markers in various types of bone and soft-tissue sarcomas. Special attention is paid to the detection of CD133, ABC transporters, nestin and aldehyde dehydrogenase that have been intensively studied both in tumour tissues and in sarcoma cell lines during the past few years. Finally, an overview is given of the possible CSC phenotypes provided by functional assays of tumourigenicity.
Asunto(s)
Biomarcadores de Tumor/análisis , Células Madre Neoplásicas/metabolismo , Sarcoma/metabolismo , Antígeno AC133 , Transportadoras de Casetes de Unión a ATP/análisis , Aldehído Deshidrogenasa/análisis , Antígenos CD/análisis , Glicoproteínas/análisis , Humanos , Proteínas de Filamentos Intermediarios/análisis , Proteínas del Tejido Nervioso/análisis , Nestina , Péptidos/análisis , Sarcoma/diagnósticoRESUMEN
Methotrexate, a structural analogue of folic acid, is one of the most frequently used chemotherapeutics, especially in haematological malignancies, various solid tumours and also inflammatory disorders. Methotrexate interferes with folate metabolism, mainly by inhibition of dihydrofolate reductase, resulting in the suppression of purine and pyrimidine precursor synthesis. The depletion of nucleic acid precursors seems to be responsible for the cytostatic, cytotoxic and differentiation effects of methotrexate. Methylation of biomolecules represents another folate-dependent pathway that is also affected by methotrexate. Furthermore, methotrexate is able to modify metabolic pathways and cellular processes independently of folate metabolism. Based on the similar structure of methotrexate and of functional groups of certain histone deacetylase inhibitors, the ability of methotrexate to inhibit histone deacetylases was predicted and consequently verified. Recently published findings also suggest that methotrexate affects glyoxalase and antioxidant systems. Although methotrexate has been used as a folate metabolism antagonist in anticancer therapy for more than 60 years, the identification of its'other molecular targets in cellular metabolism still continues.
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Antimetabolitos Antineoplásicos/farmacología , Metotrexato/farmacología , Muerte Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Ácido Fólico/metabolismo , Antagonistas del Ácido Fólico/farmacología , Inhibidores de Histona Desacetilasas/farmacología , Humanos , Metilación/efectos de los fármacos , Nucleótidos/biosíntesis , Tetrahidrofolato Deshidrogenasa/farmacologíaRESUMEN
The aim of this review is to summarize current knowledge on nestin expression in human tumors and corresponding tumor cell lines. Nestin belongs to class VI of the intermediate filaments and it is expressed primarily in mammalian nervous tissue during embryonic development. In adults, nestin occurs only in a small subset of cells and tissues. This protein has been observed in the subventricular zone of the adult mammalian brain, where neurogenesis is localized. Nestin expression has also been detected in various types of human solid tumors, as well as in the corresponding established cell lines. This article provides an up-to-date overview of tumors in which nestin has been found. Another aim of this review is to summarize recent findings on the intracellular localization of nestin in human tumor cells, especially with regard to the possible correlation between nestin expression and the malignant phenotype of transformed cells. Nestin expression in vascular endothelial cells during angiogenesis is also reviewed. Special attention is paid to the detection of nestin in cancer stem cells because this protein, together with the CD133 surface molecule, is considered to be a possible marker of cancer stem cells, especially in tumors of neuroectodermal origin.
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Proteínas de Filamentos Intermediarios/metabolismo , Neoplasias/metabolismo , Neovascularización Patológica/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Animales , Línea Celular Tumoral , Endotelio Vascular/metabolismo , Humanos , Neoplasias/patología , NestinaRESUMEN
Glioblastoma multiforme (GBM) is the most common as well as the most aggressive type of primary brain tumor of astrocytic origin in adults. GBM is characterized by a high degree of intratumoral heterogeneity both in histomorphology and genetic changes. Trisomy/polysomy of chromosome 7, monosomy of chromosome 10, EGFR gene amplification and p53 deletion have been described as the typical genetic markers for tumor classification and prediction of possible response to therapy. Our work was based on detection of these four main genetic changes both in central and peripheral parts of the tumors to evaluate possible differences in the topological incidence of these genetic markers. Chromosomal abnormalities in tumor samples from a group of 21 patients surgically treated for GBM were characterized by means of the interphase-fluorescence in situ hybridization (I-FISH) technique using sets of centromere and locus-specific DNA probes. In addition, we performed a detailed analysis of one selected tumor sample using a genomic microarray system (GenoSensor Array 300) to characterize copy number changes of specific sequences and refine results obtained by I-FISH. However, the data show no significant differences in occurrence of the described genetic markers in either part of the tumor.
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Neoplasias Encefálicas/genética , Cromosomas Humanos Par 10/genética , Receptores ErbB/genética , Glioblastoma/genética , Proteína p53 Supresora de Tumor/genética , Adulto , Anciano , Biomarcadores de Tumor/genética , Neoplasias Encefálicas/patología , Aberraciones Cromosómicas , Mapeo Cromosómico , Femenino , Amplificación de Genes , Dosificación de Gen , Marcadores Genéticos/genética , Glioblastoma/patología , Humanos , Hibridación Fluorescente in Situ , Incidencia , Cariotipificación , Masculino , Persona de Mediana Edad , Hibridación de Ácido Nucleico , Reacción en Cadena de la Polimerasa , PronósticoRESUMEN
Spectral karyotyping (SKY) represents an important tool for the investigation of the complex chromosomal rearrangements (CCRs) in many human malignancies which may be difficult to characterize by conventional banding techniques. The main goal of our work was to optimize the most important steps in the preparation of molecular cytogenetic slides for a SKY protocol. This approach consisted of optimization of both the aging procedure and protease pretreatment of the slides, with special regard given to the preservation of chromosome structure and shape, as well as to the intensity of hybridization signals. The best results were obtained with a chemical aging procedure using SSC or ethanol in combination with trypsin pretreatment applied at a higher concentration for a shorter period of pretreatment. A resulting protocol for SKY also applicable to human solid tumour cells was subsequently proposed. The practical potential of the SKY technique was demonstrated on examples of two types of human embryonal tumours--neuroblastoma and Wilms' tumour, in which some kinds of chromosomal aberrations were not detectable by means of classic cytogenetic methods.
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Neuroblastoma/genética , Neuroblastoma/patología , Manejo de Especímenes/métodos , Cariotipificación Espectral/métodos , Tumor de Wilms/genética , Tumor de Wilms/patología , Células Sanguíneas/citología , Células Sanguíneas/efectos de los fármacos , Senescencia Celular/efectos de los fármacos , Niño , Humanos , Indoles , Metafase/efectos de los fármacos , Hibridación de Ácido Nucleico , Péptido Hidrolasas/farmacologíaRESUMEN
Possible UVC-protective properties of CA, a plant phenolic compound with antioxidant activity, were investigated on human KF1 diploid fibroblast and A431 epidermoid carcinoma cell lines. Cell populations, untreated and treated by antioxidants (CA and alpha-tocopherol), were irradiated by UVC at the wavelength of 254 nm and their proliferation activity was determined by the MTT assay. The results show a strong protective effect of CA at both concentrations used (55.5 and 166.5 microM): a significant increase of proliferation activity after UVC irradiation was detected in both cell populations growing in the presence of CA in comparison with cells in DMEM only. The described protective effect of CA was more obvious in transformed cells than in normal diploid cells. This protective ability is probably based on the antioxidant and scavenging activities of CA, which seems to be more efficient than alpha-tocopherol in protection against the cytotoxic effect caused by UVC irradiation.
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Ácidos Cafeicos/farmacología , Transformación Celular Neoplásica/efectos de los fármacos , Transformación Celular Neoplásica/efectos de la radiación , Sustancias Protectoras/farmacología , Piel/efectos de los fármacos , Piel/efectos de la radiación , Rayos Ultravioleta , División Celular/efectos de los fármacos , División Celular/efectos de la radiación , Línea Celular , Transformación Celular Neoplásica/patología , Humanos , Piel/citología , Piel/patologíaRESUMEN
Micromanipulation is a strong mechanical intervention into cellular integrity and induces large changes in the fine structure of the treated cells. Human diploid skin fibroblasts (KF1 and KF2 cell lines) were chosen as an experimental model. Special hatching needles were used for defined micromanipulation interventions (deformation of plasma membrane). Changes in cytoskeletal structures were visualized by using fluorescent and confocal microscopy. The actin cytoskeleton showed a more sensitive response to micromanipulation than microtubules. Characteristic changes in microfilaments, i.e., thickenings and knot formation, were visible in treated cells fixed immediately after micromanipulation and were the result of hatching-needle pressure on the plasma membrane as well as a reaction of actin filaments localized near the plasma membrane deformation. These direct changes and also other specific alterations in the actin filament network were detectable 14 to 16 h after treatment, but they were not observed when longer reparation intervals were used.
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Citoesqueleto/ultraestructura , Micromanipulación , Piel/ultraestructura , Citoesqueleto de Actina/ultraestructura , Actinas/ultraestructura , Línea Celular , Fibroblastos/ultraestructura , Humanos , Microscopía Confocal , Microscopía Fluorescente , Microtúbulos/ultraestructuraRESUMEN
The cytoskeleton, in addition to its structural and kinetic functions, is also involved in modulating signal transfer in cell proliferation, differentiation and death. In some myeloid leukemic cell lines, the process of cell differentiation accompanied by apoptosis, can be induced by all-trans retinoic acid (ATRA). In this report, we describe the morphological changes in actin cytoskeleton, taking place during apoptosis in cells of the human leukemic HL-60 cell line. By using fluorescent microscopy, the morphology of microfilaments and the proportion of apoptotic cells in the cell populations untreated or treated with 10(-6) M ATRA were detected. Interphase HL-60 cells showed aggregations of short, thick microfilament bundles in the region between the plasma membrane and the nucleus. In comparison with both interphase and mitotic cells, the cells with apoptotic nuclear fragmentation showed a different organisation of the actin cytoskeleton. The following types of F-actin structures were observed: (i) Cells with a high number of large dots/patches of F-actin under the plasma membrane. These dots might be localised only in the part of the cell or occurred under the whole plasma membrane. This arrangement was often associated with a diffuse signal for F-actin. (ii) Cells with 3D-network of F-actin fibres through the cytoplasm between remnants of the cell nucleus. This 3D-structure probably played an important active role in the process of apoptotic bodies formation. (iii) Cells without any detectable signal for F-actin or cells with only a very low F-actin signal. Both of these showed typical apoptotic collapse of chromatin. It is concluded that the actin cytoskeleton is a dynamic structure actively involved in the executive phase of the process of apoptosis. It is suggested that the rearrangement of the microfilament network and its subsequent degradation are necessary for the main morphological changes of apoptotic cells, i.e., plasma membrane blebbing and apoptotic bodies formation.
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Actinas/metabolismo , Citoesqueleto/efectos de los fármacos , Tretinoina/farmacología , Apoptosis/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Células HL-60 , Humanos , Leucocitos/citología , Leucocitos/efectos de los fármacos , Microscopía Fluorescente , Células Mieloides/citología , Células Mieloides/efectos de los fármacos , Factores de TiempoRESUMEN
The Somatic Mutation and Recombination Test (SMART) in wing cells of Drosophila melanogaster, the Vicia faba cytogenetic tests-Sister Chromatid Exchange (SCE) and Micronucleus Test (MN), and the Müller test for gametic mutations in Arabidopsis thaliana were used for genotoxicity testing of environmental samples of pollutants from the surroundings of LACHEMA chemical factory (Brno, Czech Republic) and DEZA factory in Valasské Mezirící (Moravia, Czech Republic). Tested soil and air samples were taken from the near vicinity of both factories. The surroundings of both sites are heavy loaded by exhalation of chemicals from the factories. Chemical analyses of the 16 Polycyclic Aromatic Hydrocarbons (PAHs) according to the United States Environmental Protection Agency (US EPA) list of priority pollutants and heavy metals were performed in both soil and air samples. The Drosophila wing spot test was positive in 70.6% of the tested samples, the Vicia sister chromatid exchange test in 62.5%, and the Arabidopsis Müller test in 58.9%. The micronucleus Vicia faba test was quite insensitive in tested environmental samples. The concordance between SMART and SCE was 62.5%, between SMART and Müller test 76.5%, and between Müller test and SCE 100%. Total concordance of these three tests was 79.7%. Müller test for gametic mutation in Arabidopsis thaliana and cytogenetic SCE test in Vicia faba seem to be quite sensitive and convenient plant bioassays for assessing the mutagenic potential of environmental agents, when compared to the SMART test in Drosophila melanogaster.