RESUMO
Glia cells provide supportive functions to the central nervous system and can be compromised by environmental contaminants. The primary objective of this study was to characterize the effects of in vitro exposure to perfluorooctanoic acid, a persistent environmental contaminant and/or monocrotophos (MCP), a neurotoxic organophosphate that is rapidly metabolized, to astroglia SVG p12 cells. The endpoints evaluated include cell viability, intracellular glutamate levels as a marker of astrocyte homeostasis function, differential gene expression for selected proteins, which include inflammatory markers (tachykinin), astrocytosis (nestin), S100B, and metabolism enzymes (CYP1A1). The results from cell viability revealed significant differences from the controls at some of the concentrations tested. Also, intracellular glutamate levels were elevated at the 10-µM concentration for perfluorooctanoic acid (PFOA) as well as the 10-µM PFOA/5-µM MCP concentration. Gene expression results at 80-µM PFOA concentration revealed a significant increase in the expression of S100B, tachykinin and CYP1A1. A combination of 10-µM PFOA/20-µM MCP caused a significant decrease in the expression of tachykinin. Gene expression for MCP exposures produced a decrease at the 20-µM MCP concentration. Immunofluorescence results indicated an increase in nestin protein expression for the 20-µM concentration of MCP, which contradicted the gene expression at the same concentration tested. The results indicate that toxicity to glia cells can compromise critical glia functions and could be implicated in neurodegenerative diseases.
Assuntos
Astrócitos/efeitos dos fármacos , Caprilatos/toxicidade , Fluorocarbonos/toxicidade , Inseticidas/toxicidade , Monocrotofós/toxicidade , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocromo P-450 CYP1A1/biossíntese , Poluentes Ambientais/toxicidade , Feminino , Expressão Gênica/efeitos dos fármacos , Ácido Glutâmico/metabolismo , Homeostase/efeitos dos fármacos , Humanos , Nestina/biossíntese , Células PC12 , Gravidez , Ratos , Subunidade beta da Proteína Ligante de Cálcio S100/biossíntese , Taquicininas/biossínteseRESUMO
Cardiomyocyte migration represents a requisite event of cardiogenesis and the regenerative response of the injured adult zebrafish and neonatal rodent heart. The present study tested the hypothesis that the appearance of the intermediate filament protein nestin in neonatal rat ventricular cardiomyocytes (NNVMs) was associated in part with the acquisition of a migratory phenotype. The cotreatment of NNVMs with phorbol 12,13-dibutyrate (PDBu) and the p38α/ß mitogen-activated protein kinase inhibitor SB203580 led to the de novo synthesis of nestin. The intermediate filament protein was subsequently reorganized into a filamentous pattern and redistributed to the leading edge of elongated membrane protrusions translating to significant lengthening of NNVMs. PDBu/SB203580 treatment concomitantly promoted the reorganization of nonmuscle myosin IIB (NMIIB) located predominantly at the periphery of the plasma membrane of NNVMs to a filamentous phenotype extending to the leading edge of elongated membrane protrusions. Coimmunoprecipitation assay revealed a physical interaction between NMIIB and nestin after PDBu/SB203580 treatment of NNVMs. In wild-type and heterozygous NMIIB embryonic hearts at E11.5-E14.5 days, nestin immunoreactivity was identified in a subpopulation of cardiomyocytes elongating perpendicular to the compact myocardium, at the leading edge of nascent trabeculae and during thickening of the compact myocardium. In mutant embryonic hearts lacking NMIIB protein expression, trabeculae formation was reduced, the compact myocardium significantly thinner and nestin immunoreactivity undetectable in cardiomyocytes at E14.5 days. These data suggest that NMIIB and nestin may act in a coordinated fashion to facilitate the acquisition of a migratory phenotype in neonatal and embryonic cardiomyocytes.
Assuntos
Coração/crescimento & desenvolvimento , Proteína Quinase 14 Ativada por Mitógeno/genética , Nestina/biossíntese , Miosina não Muscular Tipo IIB/genética , Organogênese/genética , Animais , Animais Recém-Nascidos/genética , Animais Recém-Nascidos/crescimento & desenvolvimento , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Coração/efeitos dos fármacos , Ventrículos do Coração/efeitos dos fármacos , Ventrículos do Coração/crescimento & desenvolvimento , Humanos , Imidazóis/farmacologia , Miocárdio/metabolismo , Miocárdio/patologia , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Nestina/genética , Dibutirato de 12,13-Forbol/farmacologia , Piridinas/farmacologia , Ratos , Peixe-Zebra/genéticaRESUMO
AIMS: Some studies investigated the association between nestin and the overall survival (OS) of nonsmall cell lung cancer (NSCLC). However, the results were conflicted and inconclusive. Therefore, we performed this meta-analysis to determine the association between nestin and OS of NSCLC. MATERIALS AND METHODS: PubMed and EMBASE were searched to find relevant studies. The strength of the association was calculated with the hazard ratios (HRs) and respective 95% confidence intervals (CIs). RESULTS: High expression of nestin was significantly associated with OS of NSCLC (HR = 2.09; 95% CI = 1.59-2.77). In the stratified analysis by race, we found that the expression of nestin was significantly associated with OS of NSCLC in Asians (HR = 3.02; 95% CI = 1.80-5.07) and Caucasians (HR = 1.81; 95% CI = 1.21-2.71). In addition, when we limited the meta-analysis to studies that controlled for clinical parameters, a significant association between nestin and OS of NSCLC remained (HR = 2.19; 95% CI = 1.54-3.11). A sensitivity analysis showed no substantial modification of the estimates after exclusion of individual studies. CONCLUSIONS: In conclusion, this meta-analysis suggested that high expression of nestin was significantly associated with OS of NSCLC.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/mortalidade , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/mortalidade , Nestina/biossíntese , Povo Asiático/estatística & dados numéricos , Biomarcadores Tumorais/biossíntese , Biomarcadores Tumorais/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Humanos , Neoplasias Pulmonares/patologia , Prognóstico , Taxa de Sobrevida , População Branca/estatística & dados numéricosRESUMO
INTRODUCTION: The olfactory neuro-epithelium has an intrinsic capability of renewal during lifetime provided by the existence of globose and horizontal olfactory precursor cells. Additionally, mesenchymal stromal olfactory cells also support the homeostasis of the olfactory mucosa cell population. Under in vitro culture conditions with Dulbecco modified eagle/F12 medium supplemented with 10% fetal bovine serum, tissue biopsies from upper turbinate have generated an adherent population of cells expressing mainly mesenchymal stromal phenotypic markers. A closer examination of these cells has also found co-expression of olfactory precursors and ensheathing cell phenotypic markers. These results were suggestive of a unique property of olfactory mesenchymal stromal cells as potentially olfactory progenitor cells. OBJECTIVE: To study whether the expression of these proteins in mesenchymal stromal cells is modulated upon neuronal differentiation. MATERIALS AND METHODS: We observed the phenotype of olfactory stromal cells under DMEM/F12 plus 10% fetal bovine serum in comparison to cells from spheres induced by serum-free medium plus growth factors inducers of neural progenitors. RESULTS: The expression of mesenchymal stromal (CD29+, CD73+, CD90+, CD45-), horizontal basal (ICAM-1/CD54+, p63+, p75NGFr+), and ensheathing progenitor cell (nestin+, GFAP+) proteins was determined in the cultured population by flow cytometry. The determination of Oct 3/4, Sox-2, and Mash-1 transcription factors, as well as the neurotrophins BDNF, NT3, and NT4 by RT-PCR in cells, was indicative of functional heterogeneity of the olfactory mucosa tissue sample. CONCLUSIONS: Mesenchymal and olfactory precursor proteins were downregulated by serum-free medium and promoted differentiation of mesenchymal stromal cells into neurons and astroglial cells.
Introducción. El recambio celular del neuroepitelio olfatorio ocurre durante la vida del individuo gracias a precursores olfatorios. Además, las células mesenquimales del estroma también contribuyen a la homeostasis de la mucosa. Cuando un explante de una biopsia de mucosa se cultiva en un medio esencial mínimo, se genera una población predominante de células adherentes que expresan proteínas típicas de las células mesenquimales del estroma. La coexpresión de marcadores fenotípicos de precursores olfatorios y de células del recubrimiento del nervio olfatorio constituiría una propiedad única de las células mesenquimales del estroma. Objetivo. Determinar si la diferenciación celular de las células mesenquimales hacia fenotipos neurales modula la expresión de los marcadores mesenquimales característicos. Materiales y métodos. Se compararon las células aisladas de la mucosa olfatoria en un medio de cultivo con suplemento de 10 % de suero fetal bovino con esferas generadas en un medio sin suero más factores de crecimiento. Resultados. Se determinó la expresión de proteínas de las células mesenquimales del estroma (CD29+, CD73+, CD90+, CD45-), de las basales horizontales (ICAM-1/CD54+, p63+, p75NGFr+), y de las del recubrimiento del nervio olfatorio (nestin+, GFAP+) en la misma población cultivada. La determinación de Oct 3/4, Sox-2 y Mash-1, así como de las neurotrofinas BDNF, NT3 y NT4, sugirió que las células del estroma son funcionales. La expresión de las proteínas de las células mesenquimales y los precursores olfatorios, disminuyó en las células de las mesenesferas inducidas por ausencia de suero en el medio de cultivo. Conclusión. Las células mesenquimales del estroma de la mucosa olfatoria presentan una tendencia dominante hacia la diferenciación neural.
Assuntos
Células-Tronco Mesenquimais/metabolismo , Mucosa Nasal/citologia , Mucosa Olfatória/citologia , Biossíntese de Proteínas , Adipogenia , Antígenos de Diferenciação/análise , Técnicas de Cultura de Células , Diferenciação Celular , Células Cultivadas , Condrogênese , Meios de Cultura/farmacologia , Meios de Cultura Livres de Soro/farmacologia , Proteína Glial Fibrilar Ácida/biossíntese , Proteína Glial Fibrilar Ácida/genética , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Mucosa Nasal/metabolismo , Fatores de Crescimento Neural/biossíntese , Fatores de Crescimento Neural/genética , Nestina/biossíntese , Nestina/genética , Neuroglia/metabolismo , Neurônios/metabolismo , Mucosa Olfatória/metabolismo , Osteogênese , Proteínas Recombinantes/farmacologia , Esferoides Celulares , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genética , Conchas NasaisRESUMO
With the aid of extrusion-based biofabrication strategies, neural stem/progenitor cell-laden hydrogel structures can be fabricated for use in neural research. Extrusion-based strategies can be altered in order to fulfill various requirements. In this study, mouse neural progenitor cell (NE-4C) behaviors in multiple extrusion-based fabricated microenvironments were investigated. Extrusion-based bioprinted cell-laden structures and coaxially extruded core-shell cell fibers were successfully fabricated. Cell distribution and morphology were observed in different structures with scanning electron microscopy (SEM). Genes and proteins related to cell differentiation were examined using quantitative polymerase chain reaction (qPCR) and western blot (WB). The results show that compared with NE-4Cs cultured in petri dishes, the abundance of nestin was 6.28 ± 1.38 times higher in bioprinted structures and the abundances of Tuj-1 and GFAP were 3.14 ± 1.38 and 2.11 ± 0.21 times higher in cell fibers, respectively, indicating that NE-4Cs showed stronger differentiation tendency in cell fibers and weaker tendency in printed structures. This study may provide guidance in selecting fabrication strategies for use in neural research.
Assuntos
Bioimpressão/métodos , Hidrogéis/química , Nestina/biossíntese , Células-Tronco Neurais/citologia , Animais , Diferenciação Celular , Linhagem Celular , Sobrevivência Celular , Células Cultivadas , Camundongos , Microscopia Eletrônica de Varredura , Nestina/química , Impressão TridimensionalRESUMO
Hair-follicle-associated pluripotent (HAP) stem cells reside in the upper part of the bulge area of the the hair follicle. HAP stem cells are nestin-positive and keratin 15-negative and have the capacity to differentiate into various types of cells in vitro. HAP stem cells are also involved in nerve and spinal cord regeneration in mouse models. Recently, it was shown that the DNA-damage response in non-HAP hair follicle stem cells induces proteolysis of type-XVII collagen (COL17A1/BP180), which is involved in hair-follicle stem-cell maintenance. COL17A1 proteolysis stimulated hair-follicle stem-cell aging, characterized by the loss of stemness signatures and hair-follicle miniaturization associated with androgenic alopecia. In the present study, we demonstrate that HAP stem cells co-express nestin and COL17A1 in vitro and in vivo. The expression of HAP stem cell markers (nestin and SSEA1) increased after HAP stem-cell colonies were formed, then decreased after differentiation to epidermal keratinocytes. In contrast COL17A1 increased after differentiation to epidermal keratinocytes. These results suggest that COL17A1 is important in differentiation of HAP stem cells.
Assuntos
Autoantígenos/biossíntese , Diferenciação Celular , Regulação da Expressão Gênica , Folículo Piloso/metabolismo , Queratinócitos/metabolismo , Colágenos não Fibrilares/biossíntese , Células-Tronco Pluripotentes/metabolismo , Animais , Antígenos de Diferenciação/biossíntese , Folículo Piloso/citologia , Queratinócitos/citologia , Camundongos , Nestina/biossíntese , Células-Tronco Pluripotentes/citologia , Colágeno Tipo XVIIRESUMO
In the prostate, stem and progenitor cell regenerative capacities have been ascribed to both basal and luminal epithelial cells. Here, we show that a rare subset of mesenchymal cells in the prostate are epithelial-primed Nestin-expressing cells (EPNECs) that can generate self-renewing prostate organoids with bipotential capacity. Upon transplantation, these EPNECs can form prostate gland tissue grafts at the clonal level. Lineage-tracing analyses show that cells marked by Nestin or NG2 transgenic mice contribute to prostate epithelium during organogenesis. In the adult, modest contributions in repeated rounds of regression and regeneration are observed, whereas prostate epithelial cells derived from Nestin/NG2-marked cells are dramatically increased after severe irradiation-induced organ damage. These results indicate that Nestin/NG2 expression marks a novel radioresistant prostate stem cell that is active during development and displays reserve stem cell activity for tissue maintenance.
Assuntos
Antígenos/biossíntese , Células Epiteliais/metabolismo , Nestina/biossíntese , Transplante de Órgãos , Próstata/metabolismo , Próstata/transplante , Proteoglicanas/biossíntese , Lesões Experimentais por Radiação , Tolerância a Radiação , Células-Tronco/metabolismo , Animais , Antígenos/genética , Células Epiteliais/patologia , Regulação da Expressão Gênica/efeitos da radiação , Masculino , Camundongos , Camundongos Transgênicos , Nestina/genética , Próstata/patologia , Proteoglicanas/genética , Lesões Experimentais por Radiação/metabolismo , Lesões Experimentais por Radiação/patologia , Lesões Experimentais por Radiação/cirurgia , Células-Tronco/patologiaRESUMO
Nestin-expressing stromal cells (NESCs) and Schwann cells in the bone marrow (BM) play crucial roles as a niche for normal hematopoietic stem cells in mice. It has been reported that both types of cells are decreased in myeloproliferative neoplasms in patients and also in a mouse model, whereas an increase in NESCs was reported in acute myeloid leukemia. It is thus of interest whether and how these BM stromal cells are structured in myelodysplastic syndromes (MDS). Here, we focused on NESCs and glial fibrillary acidic protein (GFAP)-expressing cells in the BM of MDS patients. We found a marked increase of NESCs in MDS with fibrosis (MDS-F) at a high frequency (9/19; 47.4%), but not in MDS without fibrosis (0/26; 0%). Intriguingly, in eight of the nine (88.9%) MDS-F cases with elevated NESCs, a majority of NESCs also expressed GFAP, with an additional increase in GFAP single-positive cells. Furthermore, in seven of them, we found a prominent structure characterized by neurofilament heavy chain staining surrounded by NESCs with GFAP expression. This structure may represent peripheral nerve axons surrounded by Schwann cells, and could be relevant to the pathophysiology of MDS-F.
Assuntos
Células da Medula Óssea/metabolismo , Síndromes Mielodisplásicas/metabolismo , Nestina/biossíntese , Mielofibrose Primária/metabolismo , Células de Schwann/metabolismo , Adulto , Animais , Células da Medula Óssea/patologia , Feminino , Proteína Glial Fibrilar Ácida/biossíntese , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Síndromes Mielodisplásicas/patologia , Mielofibrose Primária/patologia , Células de Schwann/patologiaRESUMO
BACKGROUND: Trichoblastoma (TB) and basal cell carcinoma (BCC) are 2 different neoplasms composed of basaloid cells and have overlapping histopathological features. We compared the immunoexpression of CD10, T-cell death-associated gene 51 (TDAG51), cytokeratin 20 (CK20), androgen receptor (AR), insulinoma-associated protein 1 (INSM1), and nestin for the differential diagnosis of these tumors. MATERIALS AND METHODS: We assessed a total of 27 BCC and 27 TB cases, including 4 TB lesions in nevus sebaceous and 3 malignant TB lesions for CD10, TDAG51, CK20, AR, INSM1, and nestin expression. RESULTS: Staining for CK20, TDAG51, INSM1, and stromal CD10 was significantly more common in TB cases than in BCC cases ( P < .001). Epithelial CD10 and AR staining was significantly more common in BCC cases than in TB cases ( P < .001). The difference between the groups for nestin staining was not significant ( P > .05). Stromal CD10 staining was the most sensitive marker (96.3%) and INSM1 the least sensitive (55.6%) marker for TB. TDAG51 showed 100% specificity for TB. A larger number of CK20 positive cells was found in the cases associated with nevus sebaceous than in the other TBs. CONCLUSION: All the selected markers except nestin were useful for the differential diagnosis between TB and BCC. CD10 and TDAG51 were more useful than the other markers. The use of CK20 could be preferred in nevus sebaceous lesions. INSM1 was less effective in highlighting Merkel cells within the lesion than CK20.
Assuntos
Carcinoma Basocelular/diagnóstico , Doenças do Cabelo/diagnóstico , Folículo Piloso/patologia , Neoplasias Cutâneas/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/análise , Diagnóstico Diferencial , Feminino , Humanos , Queratina-20/biossíntese , Masculino , Pessoa de Meia-Idade , Neprilisina/biossíntese , Nestina/biossíntese , Receptores Androgênicos/biossíntese , Proteínas Repressoras/biossíntese , Fatores de Transcrição/biossínteseRESUMO
While histological analysis represents a powerful tool for the classification of melanocytic lesions as benign or malignant, a clear-cut distinction between a nevus and a melanoma is sometimes a challenging step of the diagnostic process. The immunohistochemical detection of tyrosinase, cardinal melanogenic enzyme during melanocytic maturation, has often been helpful in formulating a differential diagnosis due to the peculiar staining pattern in nevocytes compared with melanoma cells. Tyrosinase distribution in nevi appears to overlap with the cytoarchitectural changes observable within these lesions, that result in epidermal or superficial dermal nevocytes being larger and strongly expressing melanocytic differentiation antigens, such as tyrosinase, compared with deeper dermal nevus cells. Our study aimed to evaluate the immunohistochemical expression pattern of tyrosinase in different histological types of acquired dysplastic melanocytic nevi, including junctional, compound, and intradermal nevi. Moreover, to estimate whether in nevocytes the expression of tyrosinase was associated with their differentiation state, we investigated the expression of two recognized markers of pluripotency, CD34 and nestin. In all examined nevi, our analysis revealed a remarkable immunoreactivity for tyrosinase in junctional and superficial dermal nevocytes and a decreasing gradient of staining in dermal nevocytes, up to become negative in deeper dermis. Meanwhile, junctional and dermal nevocytes were lacking in CD34 protein. Furthermore, nestin immunostaining showed an opposite distribution compared with tyrosinase, leading us to look into the tyrosinase/nestin expression pattern in melanocytic nevus as a tool to better understand the final stages of differentiation of melanocyte precursors toward their ultimate anatomical site into the epidermis.
Assuntos
Diferenciação Celular , Melanócitos/química , Melanócitos/patologia , Monofenol Mono-Oxigenase/análise , Nestina/análise , Nevo Pigmentado/química , Nevo Pigmentado/patologia , Adolescente , Adulto , Idoso , Feminino , Humanos , Imuno-Histoquímica , Masculino , Melanócitos/metabolismo , Pessoa de Meia-Idade , Monofenol Mono-Oxigenase/biossíntese , Nestina/biossíntese , Nevo Pigmentado/metabolismo , Adulto JovemRESUMO
Vascular smooth muscle cells (SMCs), distinguished by the expression of the neuronal stem cell marker nestin, may represent stem cell-like progenitor cells in various organs including the testis. We investigated epididymal tissues of adult nestin-GFP mice, rats after Leydig cell depletion via ethane dimethane sulfonate (EDS), rats and mice during postnatal development and human tissues. By use of Clarity, a histochemical method to illustrate a three-dimensional picture, we could demonstrate nestin-GFP positive cells within the vascular network. We localized nestin in the epididymis in proliferating vascular SMCs by colocalization with both smooth muscle actin and PCNA, and it was distinct from CD31-positive endothelial cells. The same nestin localization was found in the human epididymis. However, nestin was not found in SMCs of the epididymal duct. Nestin expression is high during postnatal development of mouse and rat and down-regulated towards adulthood when testosterone levels increase. Nestin increases dramatically in rats after Leydig cell ablation with EDS and subsequently low testosterone levels. Interestingly, during this period, the expression of androgen receptor in the epididymis is low and increases until nestin reaches normal levels of adulthood. Here we show that nestin, a common marker for neuronal stem cells, is also expressed in the vasculature of the epididymis. Our results give new insights into the yet underestimated role of proliferating nestin-expressing vascular SMCs during postnatal development and repair of the epididymis.
Assuntos
Regulação da Expressão Gênica , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Nestina/biossíntese , Testosterona/deficiência , Animais , Epididimo/irrigação sanguínea , Epididimo/crescimento & desenvolvimento , Epididimo/patologia , Masculino , Camundongos , Camundongos Transgênicos , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/patologiaRESUMO
The Nestin gene encodes type VI intermediate filament and is known to be expressed in undifferentiated cells during neurogenesis and myogenesis. To regulate Nestin expression, the first or second intron enhancer is activated in a tissue-dependent manner, for example, the former in mesodermal cells and the latter in neural stem cells. Although Nestin has also been used as a differentiation marker for odontoblasts during tooth development, how Nestin expression is regulated in odontoblasts remains unclear. Therefore, this study aimed to compare the expression patterns of Nestin-GFP (green fluorescent protein) with that of endogenous Nestin in developing teeth of Nestin-EGFP (enhanced GFP) transgenic mice, in which the second intron enhancer is connected with the EGFP domain, at postnatal 7d, 3w, and 8w. Immunohistochemical and in situ hybridization analyses revealed that endogenous Nestin protein and Nestin mRNA were intensely expressed in differentiated odontoblasts, while GFP immunoreactivity, which reflects the activity of Nestin second intron enhancer-mediated transcription, was mainly observed in the subodontoblastic layer. These results indicate that the first intron enhancer may be activated in differentiated odontoblasts. Intriguingly, Nestin-GFP expression in the subodontoblastic layer was found to be restricted to the coronal pulp of molars, which is susceptible to tooth injuries. Because the subodontoblastic layer serves as a reservoir of newly differentiated odontoblast-like cells upon exogenous stimuli to dentin, our findings suggest that the original odontoblasts and regenerated odontoblast-like cells may differently regulate Nestin expression.
Assuntos
Nestina/biossíntese , Odontoblastos/metabolismo , Animais , Diferenciação Celular , Proteínas de Fluorescência Verde/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Nestina/genética , Odontoblastos/citologia , RNA Mensageiro/biossíntese , RNA Mensageiro/genéticaRESUMO
PURPOSE: Nestin, a member of the intermediate filament protein family, has been described as a putative cancer stem cell marker (CSC) in uveal melanoma and poor prognostic factor in a variety of tumours, including cutaneous melanoma. In this study, we examined the expression of nestin in primary (PUM) and metastatic uveal melanoma (MUM) samples, and correlated the findings with histological, clinical and survival data. METHODS: Nestin expression was assessed by immunohistochemistry in 141 PUM and 26 MUM samples; 11 PUM cases were matched with their corresponding metastases. The percentage of tumour cells expressing nestin was scored by three independent observers. Statistical analysis of all data was performed with SPSS. RESULTS: Nestin expression was identified in both the cytoplasm and membrane of UM cells. Increased expression of nestin in PUM samples was associated with known poor prognostic parameters, including epithelioid cell morphology (p < 0.001), closed loops (p = 0.001), higher mitotic count (p < 0.001), monosomy 3 (p = 0.007) and chromosome 8q gain (p < 0.001). Primary uveal melanoma (PUM) with nestin expression levels above a cut-off value of 10% [as determined by receiver operating characteristic (ROC) analysis] was associated with a significantly reduced survival time (Log-rank, p = 0.002). In MUM, a higher percentage of nestin-positive tumour cells combined with poor prognostic markers in the PUM led to a shorter survival time following the development of metastases. CONCLUSION: In conclusion, increased nestin expression in PUM is a predictor of a tumour phenotype associated with metastatic progression and reduced survival time at onset of metastasis.
Assuntos
Neoplasias Hepáticas/secundário , Melanoma/metabolismo , Estadiamento de Neoplasias , Nestina/biossíntese , Neoplasias Uveais/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/biossíntese , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/metabolismo , Masculino , Melanoma/diagnóstico , Melanoma/secundário , Pessoa de Meia-Idade , Metástase Neoplásica , Prognóstico , Neoplasias Uveais/diagnóstico , Neoplasias Uveais/secundário , Adulto JovemRESUMO
Dilated cardiomyopathy (DCM), as one of the common cardiomyopathies, is a disease of the heart muscle; however, the etiology and pathogenesis of DCM were still poorly understood. Nestin has been reported a special marker of stem/progenitor cells in various tissues, and the tissue resident Nestin+ cells could promote the wound healing and tissue remodeling. However, it remains unclear whether Nestin+ cells participate in the protection of cardiomyocytes during the pathogenesis of DCM. Here the model of mice DCM was induced by doxorubicin (DOX) intraperitoneal injection and observed heart failure and ventricular enlargement via echocardiography and histologic analysis, respectively. During DCM pathogenesis, the number of Nestin+ cells showed a significant peak on day 6 after DOX treatment, which then gradually decreases to lower than normal levels after day 30 in the total population of the heart. Furthermore, we found that the isolated increased heart-derived Nestin+ cells are mesenchymal property and could protect DOX-induced HL-1 cells toxicity in vitro by promoting their proliferation and inhibiting their apoptosis. Collectively, our results showed that Nestin+ cells increased during DCM pathogenesis and played an important role in protecting against the DOX-induced HL-1 cells loss via regulating proliferation and apoptosis. Thus, the loss of Nestin+ cells might be an etiology to DCM pathogenesis, and these cells could be a promising candidate cell source for study and treatment of DCM patients.
Assuntos
Apoptose , Cardiomiopatia Dilatada/genética , Regulação da Expressão Gênica , Ventrículos do Coração/metabolismo , Nestina/genética , RNA/genética , Função Ventricular Esquerda/fisiologia , Animais , Cardiomiopatia Dilatada/patologia , Cardiomiopatia Dilatada/fisiopatologia , Células Cultivadas , Modelos Animais de Doenças , Doxorrubicina/toxicidade , Ecocardiografia , Citometria de Fluxo , Ventrículos do Coração/patologia , Ventrículos do Coração/fisiopatologia , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Miócitos Cardíacos/patologia , Nestina/biossíntese , Reação em Cadeia da Polimerase em Tempo RealRESUMO
AIM: To investigate the expression of cancer stem cell markers in meningiomas. MATERIAL AND METHODS: CD133, Nestin and Sox2 expression levels in 35 paraffin-embedded meningioma tissue samples were assessed using immunohistochemistry. RESULTS: In this study, five cases were atypical (WHO Grade II), two were anaplastic (WHO Grade III), and 28 were benign (WHO Grade I). Among atypical and anaplastic meningiomas, all were positive for Nestin and CD133, and 4 were positive for Sox2. Of the 28 benign meningiomas, 23 were positive for Nestin, 11 were positive for CD133, and none were positive for Sox2. In addition, Nestin and CD133 were expressed at significantly higher levels in the non-benign group than in the benign group. CONCLUSION: Nestin, CD133 and Sox2 expression levels may be correlated with the WHO pathological grade. Specifically, more aggressive meningiomas are characterized by higher positivity rates and higher levels of Nestin, CD133 and Sox2 expression in positive cells.
Assuntos
Antígeno AC133/biossíntese , Biomarcadores Tumorais/análise , Neoplasias Meníngeas/patologia , Meningioma/patologia , Nestina/biossíntese , Fatores de Transcrição SOXB1/biossíntese , Adulto , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Morphine is used as an analgesic although it causes important secondary effects. These effects are triggered by several mechanisms leading to the dysregulation of gene expression. Here we aimed to study these alterations on neural stem cells (NSC) during CNS development. METHODS: AB strain and tg nestin:GFP zebrafish embryos, zebrafish primary neuron culture and mouse embryonic stem cells were used to assess the effect of morphine by qPCR, time lapse microscopy and western blot. ChIP-qPCR and bisulfite conversion assay were performed to determine the changes exerted by morphine in a Nestin candidate enhancer. RESULTS: Morphine increases GFP in nestin:GFP embryos and overexpresses the NSC marker Nestin. Morphine also exerts a hyperacetylation effect on H3K27 and decreases DNA methylation within a region located 18 Kb upstream nestin transcription starting site. Here, a binding site for the transcription factor complex Sox2/Oct4/Nanog was predicted. These factors are also upregulated by morphine. Besides, morphine increases the histone acetyl transferase p300. The inhibition of p300 activity decreases Nestin. CONCLUSIONS: Morphine facilitates Nestin increase by several mechanisms which include hyperacetylation of H3K27, decreased DNA methylation and the overexpression of the transcription factors sox2, oct4 and nanog. It has also been demonstrated that nestin levels depend on p300 activity. The facilitated Nestin expression delays the normal differentiation of neural stem cells. GENERAL SIGNIFICANCE: The present work provides novel evidence of the effects induced by morphine in the normal differentiation of NSCs, altering Nestin through changes on p300, H3K27ac, DNA methylation and Oct4, Sox2, and Nanog.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Morfina/farmacologia , Nestina/biossíntese , Células-Tronco Neurais/efeitos dos fármacos , Neurogênese/efeitos dos fármacos , Proteínas de Peixe-Zebra , Acetilação/efeitos dos fármacos , Animais , Animais Geneticamente Modificados , Sítios de Ligação , Ilhas de CpG/efeitos dos fármacos , Metilação de DNA/efeitos dos fármacos , Proteína p300 Associada a E1A/fisiologia , Embrião não Mamífero/efeitos dos fármacos , Genes Reporter , Histonas/metabolismo , Humanos , Camundongos , Naloxona/farmacologia , Proteína Homeobox Nanog/biossíntese , Proteína Homeobox Nanog/genética , Nestina/genética , Células-Tronco Neurais/metabolismo , Fator 3 de Transcrição de Octâmero/biossíntese , Fator 3 de Transcrição de Octâmero/genética , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Fatores de Transcrição SOX/biossíntese , Fatores de Transcrição SOX/genética , Regulação para Cima/efeitos dos fármacos , Proteínas de Peixe-Zebra/biossíntese , Proteínas de Peixe-Zebra/genéticaRESUMO
Astrocytes, the most abundant type of glial cells in the brain, play critical roles in supporting neuronal development and brain function. Although astrocytes have been frequently detected in brain tumors, including medulloblastoma (MB), their functions in tumorigenesis are not clear. Here, we demonstrate that astrocytes are essential components of the MB tumor microenvironment. Tumor-associated astrocytes (TAA) secrete the ligand sonic hedgehog (Shh), which is required for maintaining MB cell proliferation despite the absence of its primary receptor Patched-1 (Ptch1). Shh drives expression of Nestin in MB cells through a smoothened-dependent, but Gli1-independent mechanism. Ablation of TAA dramatically suppresses Nestin expression and blocks tumor growth. These findings demonstrate an indispensable role for astrocytes in MB tumorigenesis and reveal a novel Ptch1-independent Shh pathway involved in MB progression. Cancer Res; 77(23); 6692-703. ©2017 AACR.
Assuntos
Astrócitos/metabolismo , Carcinogênese/patologia , Neoplasias Cerebelares/patologia , Proteínas Hedgehog/metabolismo , Meduloblastoma/patologia , Animais , Proliferação de Células/fisiologia , Células Cultivadas , Camundongos , Camundongos Transgênicos , Nestina/biossíntese , Receptor Patched-1/metabolismo , Receptor Smoothened/metabolismo , Microambiente Tumoral/fisiologia , Proteína GLI1 em Dedos de Zinco/metabolismoRESUMO
The aim of the present study was to investigate changes occurring in the number of beta cells, as well as the expressions of Ngn-3, nestin and Pdx-1 of pancreatic progenitor cells in the pancreas of experimentally-induced adult diabetic rats and to determine the effect of orally-administered lycopene on these changes. Following the administration of 50 mg/kg streptozotocin to rats, four groups of animals were established: control + corn oil, control + lycopene, diabetic + corn oil and diabetic + lycopene. The animals in the control + lycopene and diabetic + lycopene groups received 4 mg/kg lycopene for a period of four weeks. The expressions of insulin, Ngn-3, nestin, and Pdx-1 were determined through immunohistochemistry in sections taken from pancreas tissue samples at the end of the experiment. The number of insulin-positive cells was found to be significantly low in the diabetic groups compared to the control groups. In addition, the presence of Ngn-3 and nestin-positive cells within the exocrine pancreas surrounding the islands was noted in the diabetic groups. Lycopene, in general did not have any effect in any of the parameters analyzed in the present study. It is suggested that these cells would function as stem cells to replace the lost beta-cell population. It is also suggested that it is possible to demonstrate the antioxidant effects of lycopene in the pancreas of diabetic rats by increasing the dose and duration of lycopene administration. Anat Rec, 300:2200-2207, 2017. © 2017 Wiley Periodicals, Inc.
Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/biossíntese , Carotenoides/farmacologia , Diabetes Mellitus Experimental/metabolismo , Proteínas de Homeodomínio/biossíntese , Proteínas do Tecido Nervoso/biossíntese , Nestina/biossíntese , Pâncreas/metabolismo , Transativadores/biossíntese , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Carotenoides/uso terapêutico , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/genética , Feminino , Proteínas de Homeodomínio/genética , Licopeno , Proteínas do Tecido Nervoso/genética , Nestina/genética , Pâncreas/efeitos dos fármacos , Ratos , Ratos Wistar , Transativadores/genéticaRESUMO
BACKGROUND: Diabetic kidney disease (DKD) is one of the most debilitating complications of type 2 diabetes. Recent evidence suggests chronic inflammation to be one of the causal factors of DKD. The mechanisms entailed are not completely elucidated except that a variety of cytokines play a major role in this process. High mobility group box 1 (HMGB1) is a pro-inflammatory toll-like receptor-4 (TLR4)-binding cytokine that is involved in inflammation-associated gene expression. This investigation was designed to assess the involvement of HMGB1, TLR-4, and nuclear factor (NF)-κB in the development of DKD and to evaluate that whether blocking HMGB1 by its natural inhibitor Glycyrrhizin (GLC) can reduce the progression of the disease. METHODS: Studies were carried out in 8-10-weeks old Zucker diabetic fatty (ZDF) and lean, age- and gender-matched rats. At 10 weeks of age, ZDF rats as compared to controls, showed hyperglycemia, without proteinuria. After 8-10 weeks of the development of diabetes, ZDF animals that showed proteinuria were treated with GLC for 4 weeks. In addition, normal rat kidney (NRK-52E) cells with epithelial-like morphology were comparatively treated with GLC under hyperglycemic condition in vitro. RESULTS: Substantial increase in the expression of HMGB1, TLR4, and NF-κB in vivo and in vitro under hyperglycemic conditions was observed as compared to normoglycemic conditions. The overexpression of HMGB1, TLR4, NF-κB, and glomerular injury marker nestin was significantly ameliorated by GLC administration. CONCLUSION: Our findings suggest that hyperglycemia-induced HMGB1 activation in ZDF rats may contribute to the progression of DKD.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Nefropatias Diabéticas/tratamento farmacológico , Ácido Glicirrízico/uso terapêutico , Nefrite/tratamento farmacológico , Animais , Linhagem Celular , Nefropatias Diabéticas/complicações , Nefropatias Diabéticas/fisiopatologia , Proteína HMGB1/biossíntese , Hiperglicemia/complicações , Inflamassomos/efeitos dos fármacos , Rim/citologia , Rim/patologia , Rim/fisiopatologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , NF-kappa B/biossíntese , Nefrite/etiologia , Nestina/biossíntese , Nestina/sangue , Proteinúria/etiologia , Ratos , Ratos Zucker , Receptor 4 Toll-Like/biossínteseRESUMO
Adipose tissue-derived stem cells (ASCs) can be obtained from different adipose tissue sources within the body. It is an abundant cell pool, easily accessible, suitable for cultivation and expansion in vitro and preparation for therapeutic approaches. Amongst these therapeutic approaches are tissue engineering and nervous system disorders such as spinal cord injuries. For such treatment, ASCs have to be reliably differentiated in to the neuronal direction. Therefore, we investigated the neural differentiation potential of ASCs using protocols with neurogenic inductors such as valproic acid and forskolin, while dog brain tissue served as control. Morphological changes could already be noticed 1 h after neuronal induction. Gene expression analysis revealed that the neuronal markers nestin and ßIII-tubulin as well as MAP2 were expressed after induction of neuronal differentiation. Additionally, the expression of the neurotrophic factors NGF, BDNF and GDNF was determined. Some of the neuronal markers and neurotrophic factors were already expressed in undifferentiated cells. Our findings point out that ASCs can reliably be differentiated into the neuronal lineage; therefore, these cells are a suitable cell source for cell transplantation in disorders of the central nervous system. Follow-up studies would show the clinical benefit of these cells after transplantation.