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1.
J Neurosci Res ; 93(12): 1891-902, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26356851

RESUMEN

miR-18a represses angiogenesis and tumor evasion by weakening vascular endothelial growth factor and transforming growth factor-ß signaling to prolong the survival of glioma patients, although it is thought to be an oncogene. This study investigates the potential effects of miR-18a on the permeability of the blood-tumor barrier (BTB) and its possible molecular mechanisms. An in vitro BTB model was successfully established. The endogenous expression of miR-18a in glioma vascular endothelial cells (GECs) was significantly lower than that in normal vascular ECs, and the overexpression of miR-18a significantly increased the permeability of the BTB as well as downregulating the mRNA and protein expressions of tight junction-related proteins zonula occluden-1 (ZO-1), claudin-5, and occludin in GECs. Dual luciferase reporter assays revealed that miR-18a bound to the 3'-untranslated region (3'UTR) of myocyte enhancer factor 2D (MEF2D). The overexpression of both miR-18a and MEF2D with the 3'UTR significantly weakened the effect caused by miR-18a of decreasing the mRNA and protein expressions of ZO-1, claudin-5 and occludin and of increasing the permeability of the BTB. Chromatin immunoprecipitation showed that MEF2D could directly bind to KLF4 promoter. This study shows that miR-18a targets and negatively regulates MEF2D, which further regulates tight junction-related proteins ZO-1, claudin-5, and occludin through transactivation of KLF4 and, finally, changes the permeability of the BTB. MiR-18a should garner growing attention because it might serve as a potential target in opening the BTB and providing a new strategy for the treatment of gliomas.


Asunto(s)
Regulación hacia Abajo/fisiología , Células Epiteliales/metabolismo , Factores de Transcripción de Tipo Kruppel/metabolismo , Factores de Transcripción MEF2/metabolismo , MicroARNs/metabolismo , Proteínas de la Zonula Occludens/metabolismo , Barrera Hematoencefálica/citología , Permeabilidad Capilar/fisiología , Línea Celular Transformada , Inmunoprecipitación de Cromatina , Claudina-5/metabolismo , Glioma/patología , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Peroxidasa de Rábano Silvestre/metabolismo , Humanos , Factor 4 Similar a Kruppel , MicroARNs/genética , Ocludina/metabolismo , Permeabilidad , ARN Mensajero/metabolismo , Transfección , Proteínas de la Zonula Occludens/genética , Proteína de la Zonula Occludens-1/metabolismo
2.
J Neurosci Res ; 92(6): 703-13, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24523141

RESUMEN

This study aims to determine the effects of vascular endothelial growth factor (VEGF), papaverine (PA), and the combination of VEGF and PA on the permeability of the blood-tumor barrier (BTB) and to determine possible molecular mechanisms contributing to the effects. In the rat C6 glioma model, the extravasation of Evans blue (EB) through the BTB was increased significantly by VEGF and PA. VEGF-induced and PA-induced increase of EB extravasation was further increased after combining VEGF with PA infusion. Transmission electron microscopy (TEM) showed that the combination of VEGF and PA not only opened tight junctions (TJ) dramatically but increased the presence of pinocytotic vesicles of brain microvascular endothelial cells (BMECs) significantly. Meanwhile, the downregulation of the TJ-associated proteins occludin and claudin-5 and the upregulation of the caveolae structure proteins caveolin-1 and caveolin-2 caused by the combination of VEGF and PA were observed by Western blot and immunohistochemistry, which were more remarkable than those by the two strategies separately. In addition, after VEGF and PA infusion, the results of radioimmunoassay, Western blot, and enzyme-linked immunosorbent assay (ELISA) revealed a significant increase in expression levels of cGMP and protein kinase G-1 (PKG-1) and the activation of nuclear factor-κB (NF-κB) p65. This study demonstrates that combination of VEGF and PA can increase the permeability of the BTB by a paracellular pathway (downregulation of occludin and claudin-5) and a transcellular pathway (upregulation of caveolin-1 and caveolin-2) and that the cGMP/PKG/NF-κB signal pathway might be involved in the modulation process.


Asunto(s)
Permeabilidad Capilar/efectos de los fármacos , Neoplasias Experimentales/irrigación sanguínea , Neoplasias Experimentales/patología , Papaverina/farmacología , Factor A de Crecimiento Endotelial Vascular/farmacología , Animales , Western Blotting , Neoplasias Encefálicas/irrigación sanguínea , Neoplasias Encefálicas/patología , Línea Celular Tumoral , Ensayo de Inmunoadsorción Enzimática , Femenino , Glioma/irrigación sanguínea , Glioma/patología , Inmunohistoquímica , Microscopía Electrónica de Transmisión , Inhibidores de Fosfodiesterasa/farmacología , Pinocitosis/efectos de los fármacos , Radioinmunoensayo , Ratas , Ratas Wistar , Uniones Estrechas/efectos de los fármacos , Uniones Estrechas/ultraestructura
3.
Yao Xue Xue Bao ; 44(8): 911-4, 2009 Aug.
Artículo en Zh | MEDLINE | ID: mdl-20055161

RESUMEN

Long circulating paclitaxel microemulsions (TXL-M) were prepared and its anticancer effect was evaluated in metronomic chemotherapy of cancer using animal tumor models. In TXL-M, paclitaxel was dissolved in vitamin E and polyethylene glycol-distearoylphosphatidylethanolamine (PEG-DSPE) was used as surfactant. The shape and particle size distribution of TXL-M were evaluated using an electronic microscope and a laser size scanner. The toxicity comparisons of TXL-M and paclitaxel were conducted using mice. Its anticancer effect and long circulation were evaluated using animal tumor model in C57BL/6 mice. The average diameter of TXL-M was (98.6 +/- 11.2) nm and its zeta potential was (-32.4 +/- 6.8) mV. Compared with paclitaxel, TXL-M showed lower toxicity. When used in metronomic chemotherapy of cancer, TXL-M showed longer circulation time in the blood and greater anticancer effect than paclitaxel. Thus, TXL-M is a better candidate for metronomic chemotherapy of cancer than paclitaxel injection.


Asunto(s)
Antineoplásicos Fitogénicos/toxicidad , Antineoplásicos Fitogénicos/uso terapéutico , Paclitaxel/toxicidad , Paclitaxel/uso terapéutico , Animales , Antineoplásicos Fitogénicos/administración & dosificación , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Paclitaxel/administración & dosificación , Células Tumorales Cultivadas
4.
Cell Signal ; 30: 142-153, 2017 01.
Artículo en Inglés | MEDLINE | ID: mdl-27693218

RESUMEN

Malignant glioma is undoubtedly the most vascularized tumor of central nervous system. Angiogenesis, playing a predominant role in tumor progression, is widely considered as a key point of tumor treatment. The aim of this study was to investigate the potential effects of miR-383 on proliferation, migration, tube formation and angiogenesis of glioma-exposed endothelial cells (GECs) in vitro and to further elucidate its possible molecular mechanisms. The expression of miR-383 in GECs was significantly downregulated compared with that in normal endothelial cells (ECs). Overexpression of miR-383 dramatically inhibited the proliferation, migration, tube formation and spheroid-based angiogenesis of GECs in vitro. Dual-luciferase reporter results demonstrated vascular endothelial growth factor (VEGF) is a target gene of miR-383. Furthermore, overexpression or silencing of either miR-383 or VEGF was performed simultaneously to further clarify that miR-383 inhibited proliferation, migration and angiogenesis of GECs in vitro by targeting VEGF. Finally, VEGF/VEGFR2-mediated FAK and Src signaling pathways might contribute to anti-angiogenesis of GECs. In conclusion, our present study indicated that miR-383 inhibits proliferation, migration and angiogenesis of GECs in vitro via VEGF/VEGFR2-mediated FAK and Src signaling pathways, which would draw growing attention to miR-383c as a potential therapeutical target of glioma.


Asunto(s)
Movimiento Celular/genética , Células Endoteliales/metabolismo , Proteína-Tirosina Quinasas de Adhesión Focal/metabolismo , Glioma/irrigación sanguínea , Glioma/patología , MicroARNs/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Familia-src Quinasas/metabolismo , Secuencia de Bases , Neoplasias Encefálicas/irrigación sanguínea , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patología , Línea Celular Tumoral , Proliferación Celular , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Glioma/genética , Humanos , MicroARNs/genética , Neovascularización Patológica/genética , Fosforilación , Transducción de Señal , Transfección , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismo
5.
Zhongguo Zhong Yao Za Zhi ; 31(8): 672-5, 2006 Apr.
Artículo en Zh | MEDLINE | ID: mdl-16830829

RESUMEN

OBJECTIVE: To study the analgesic, antipyretic and anti-inflammatory effect of Bailian Caogen granule. METHOD: The antipyretic effects of Bailian Caogen granule was evaluated in rabbit fever model induced by peptone. The analgesic effect of the drug was studied with pain model of mice induced by acetic acid and hot plate, The severity of oedema in inflamed animal was observed to study the anti-inflammatory effects of Bailian Caogen granule. RESULT: Bailian Caogen granule could obviously inhibit the fever of rabbits. The number of writhing induced by acetic acid was reduced and the pain threshold of mice was increased by Bailian Caogen granule. Bailian Caogen granule also had anti-inflammatory activity against xylene-induced mouse ear swelling and carrageenin-induced rat paw edema. CONCLUSION: Bailian Caogen granule has significant analgesic, antipyretic and anti-inflammatory activities.


Asunto(s)
Analgésicos no Narcóticos/farmacología , Medicamentos Herbarios Chinos/farmacología , Plantas Medicinales/química , Ácido Acético , Animales , Temperatura Corporal/efectos de los fármacos , Coptis/química , Combinación de Medicamentos , Edema/patología , Edema/prevención & control , Femenino , Fiebre/fisiopatología , Fiebre/prevención & control , Glycyrrhiza uralensis/química , Calor , Hiperalgesia/etiología , Hiperalgesia/fisiopatología , Hiperalgesia/prevención & control , Masculino , Ratones , Dolor/inducido químicamente , Dolor/fisiopatología , Dolor/prevención & control , Umbral del Dolor/efectos de los fármacos , Phellodendron/química , Pueraria/química , Conejos , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley
6.
CNS Neurosci Ther ; 21(1): 40-51, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25230316

RESUMEN

BACKGROUND AND AIMS: Great interest persists in useful therapeutic targets in glioblastoma (GBM). Deregulation of microRNAs (miRNAs) expression has been associated with cancer formation through alterations in gene targets. In this study, we reported the role of miR-101 in human glioblastoma stem cells (GSCs) and the potential mechanisms. METHODS AND RESULTS: Quantitative real-time PCR showed that miR-101 expression was decreased in GSCs. Overexpression of miR-101 reduced the proliferation, migration, invasion, and promoted apoptosis of GSCs. One direct target of miR-101, the transcription factor Kruppel-like factor 6 (KLF6), was identified using the Dual-Luciferase Reporter Assay System, which mediated the tumor suppressor activity of miR-101. This process was coincided with the reduced expression of Chitinase-3-like protein 1 (CHI3L1) whose promoter could be bound with and be promoted by KLF6 demonstrated by luciferase assays and chromatin immunoprecipitation assays. The downregulation of CHI3L1 led to the inactivation of MEK1/2 and PI3K signal pathways. Furthermore, nude mice carrying the tumors of overexpressed miR-101 combined with knockdown of KLF6 produced the smallest tumors and showed the highest survival rate. CONCLUSIONS: Our findings provided a comprehensive analysis of miR-101 and further defining it as a potential therapeutic candidate for GBM.


Asunto(s)
Glioblastoma/fisiopatología , Factores de Transcripción de Tipo Kruppel/metabolismo , MicroARNs/metabolismo , Células Madre Neoplásicas/fisiología , Proteínas Proto-Oncogénicas/metabolismo , Adipoquinas/genética , Adipoquinas/metabolismo , Animales , Apoptosis/fisiología , Movimiento Celular/fisiología , Proliferación Celular/fisiología , Proteína 1 Similar a Quitinasa-3 , Células HEK293 , Humanos , Factor 6 Similar a Kruppel , Factores de Transcripción de Tipo Kruppel/genética , Lectinas/genética , Lectinas/metabolismo , MAP Quinasa Quinasa 1/metabolismo , MAP Quinasa Quinasa 2/metabolismo , Ratones Mutantes Neurológicos , Invasividad Neoplásica/fisiopatología , Trasplante de Neoplasias , Fosfatidilinositol 3-Quinasas/metabolismo , Regiones Promotoras Genéticas , Proteínas Proto-Oncogénicas/genética , Transducción de Señal
7.
Cell Signal ; 27(1): 156-67, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25452107

RESUMEN

The purposes of this study were to investigate the possible molecular mechanisms of miR-18a regulating the permeability of blood-tumor barrier (BTB) via down-regulated expression and distribution of runt-related transcription factor 1 (RUNX1). An in vitro BTB model was established with hCMEC/D3 cells and U87MG cells to obtain glioma vascular endothelial cells (GECs). The endogenous expressions of miR-18a and RUNX1 were converse in GECs. The overexpression of miR-18a significantly impaired the integrity and increased the permeability of BTB, which respectively were detected by TEER and HRP flux assays, accompanied by down-regulated mRNA and protein expressions and distributions of ZO-1, occludin and claudin-5 in GECs. Dual-luciferase reporter assay was carried out and revealed RUNX1 is a target gene of miR-18a. Meanwhile, mRNA and protein expressions and distribution of RUNX1 were downregulated by miR-18a. Most important, miR-18a and RUNX1 could reversely regulate the permeability of BTB as well as the expressions and distributions of ZO-1, occludin and claudin-5. Finally, chromatin immunoprecipitation verified that RUNX1 interacted with "TGGGGT" DNA sequence in promoter region of ZO-1, occludin and claudin-5 respectively. Taken together, our present study indicated that miR-18a increased the permeability of BTB via RUNX1 mediated down-regulation of tight junction related proteins ZO-1, occludin and claudin-5, which would attract more attention to miR-18a and RUNX1 as potential targets of drug delivery across BTB and provide novel strategies for glioma treatment.


Asunto(s)
Neoplasias Encefálicas/genética , Claudina-5/metabolismo , Subunidad alfa 2 del Factor de Unión al Sitio Principal/metabolismo , Regulación hacia Abajo/genética , MicroARNs/metabolismo , Ocludina/metabolismo , Proteína de la Zonula Occludens-1/metabolismo , Secuencia de Bases , Neoplasias Encefálicas/patología , Línea Celular Tumoral , Inmunoprecipitación de Cromatina , Subunidad alfa 2 del Factor de Unión al Sitio Principal/genética , Células Endoteliales/metabolismo , Regulación Neoplásica de la Expresión Génica , Glioma/genética , Glioma/patología , Humanos , MicroARNs/genética , Datos de Secuencia Molecular , Permeabilidad , Regiones Promotoras Genéticas/genética , Unión Proteica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Uniones Estrechas/metabolismo
8.
J Mol Neurosci ; 44(2): 122-9, 2011 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-21193965

RESUMEN

The first goal of this study was to determine the effect of vascular endothelial growth factor (VEGF) on permeability of the blood-tumor barrier (BTB). The second goal was to determine possible cellular mechanisms by which VEGF increases permeability of the BTB. In the rat C6 glioma model, the permeability of the BTB was significantly increased after VEGF injection at dose of 0.05 ng/g and reached its peak at 45 min. Meanwhile, we observed that the density of pinocytotic vesicles of brain microvascular endothelial cells (BMECs) in the BTB increased dramatically by transmission electron microscopy. The immunohistochemistry and western blot analysis revealed that the expression level of caveolae structure proteins caveolin-1 and caveolin-2 in BMECs was increased after VEGF injection, peaked at 45 min, and then decreased to the untreated level. The time peak of expression level of caveolin-1 and caveolin-2 was identical with the peak time of permeability of the BTB and the density of pinocytotic vesicles. All of these results strongly indicated that VEGF increased permeability of the BTB caused by enhancement of the density of pinocytotic vesicles, and the molecular mechanism might be associated with upregulated expression of caveolin-1 and caveolin-2.


Asunto(s)
Neoplasias Encefálicas/patología , Caveolas/metabolismo , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/fisiología , Glioma/patología , Transcitosis/efectos de los fármacos , Factor A de Crecimiento Endotelial Vascular/farmacología , Animales , Barrera Hematoencefálica/efectos de los fármacos , Barrera Hematoencefálica/fisiología , Caveolas/ultraestructura , Caveolina 1/metabolismo , Caveolina 2/metabolismo , Línea Celular Tumoral , Femenino , Humanos , Permeabilidad , Distribución Aleatoria , Ratas , Ratas Wistar , Transcitosis/fisiología
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