RESUMEN
Nerve growth factor (NGF) is mainly secreted by the neuroglia cells, which can exert biological effect through its receptors on the specific target cell surface. NGF is closely related to neurocyte growth, differentiation and apoptosis. As a neurotropic virus, HSV-1 an easily lead to neurocyte, neuroglia cells death or apoptosis. In this study, the U251 human glioma cells were chosen as target cells to study the change of NGF and its receptors in the apoptosis process of HSV-1 infection. Our results showed that U251 cells were permissive to HSV-1 replication. In the apoptosis process of HSV-1 infected U251 cells, the expression of both NGF and P75NTR increased and then decreased, while the expression of TrkA decreased gradually. These result indicated that HSV-1 was able to induce the abnormal expression of NGF and its receptors in U251 cells.
Asunto(s)
Humanos , Apoptosis , Línea Celular Tumoral , Expresión Génica , Glioma , Genética , Metabolismo , Virología , Herpes Simple , Genética , Metabolismo , Virología , Herpesvirus Humano 1 , Genética , Fisiología , Factor de Crecimiento Nervioso , Genética , Metabolismo , Receptor de Factor de Crecimiento Nervioso , Genética , Metabolismo , Receptor trkA , Genética , Metabolismo , Replicación ViralRESUMEN
During the infection of host cells, IE2 protein is one of the first and most abundantly expressed products of HCMV genome, which plays an important role in the controlling of cell cycle and apoptosis. But the correlation between expression level and anti-apoptotic activity of IE2 protein is still not clear. In this study, we successfully established a HCMV IE2 protein expression cell line that was controlled by Tet-On system. The effect of IE2 protein on cell apoptosis and the expression of p53 was detected under different condition of induction. Our results showed that the IE2 protein could inhibit cell apoptosis induced by TNF-alpha. Additionally, the anti-apoptotic activity of IE2 protein seemed to be relevant to its expression level. However, we failed to detect any difference of p53 expression between the IE2 protein expression and non-expression cells. These data indicated that the IE2 protein might inhibit cell apoptosis through regulating different signal pathways.
Asunto(s)
Humanos , Antibacterianos , Farmacología , Apoptosis , Genética , Doxiciclina , Farmacología , Regulación de la Expresión Génica , Genética , Células HeLa , Proteínas Inmediatas-Precoces , Genética , Metabolismo , Plásmidos , Genética , Transactivadores , Genética , Metabolismo , Proteína p53 Supresora de Tumor , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To explore the change of endogenic nerve growth factor (NGF) expression in human glioma cells infected with human cytomegalovirus (HCMV).</p><p><b>METHODS</b>U251 cells were cultured in RPMI 1640 culture medium and infected with HCMV AD169 strain in vitro to establish a cell model of viral infection. Morphologic changes of U251 cells were observed under inverted microscope before and after infection with HCMV. Expression of NGF gene and protein of cells was detected by RT-PCR and Western blotting before and after infection with HCMV.</p><p><b>RESULTS</b>The cytopathic effects of HCMV-infected cells appeared on day 5 after infection. However, differential NGF expression was evident on day 7. NGF expression was decreased significantly in U251 cells on day 7 after infection in comparison with control group (P < 0.05).</p><p><b>CONCLUSION</b>HCMV can down-regulate endogenous NGF levels in human glioma cell line U251.</p>
Asunto(s)
Humanos , Actinas , Metabolismo , Línea Celular Tumoral , Citomegalovirus , Fisiología , Infecciones por Citomegalovirus , Patología , Técnica del Anticuerpo Fluorescente , Regulación Neoplásica de la Expresión Génica , Glioma , Genética , Patología , Virología , Modelos Biológicos , Factor de Crecimiento Nervioso , Genética , Metabolismo , ARN Mensajero , Genética , Metabolismo , Reproducibilidad de los Resultados , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Human herpes simplex virus 1 (HSV-1) causes facial,ocular,and encephalitic disease and is associated with latent infection and cancer.Here,we developed a means of studying the pathogenesis of HSV-1 infection at the protein level by using the SELDI Protein Chip to detect changes of protein expression in Vero cells cultured in vitro.After infection with HSV-1 and culture for 12,24 or 48 h,cells were harvested and lysed.IMAC3 arrays were applied to SELDI-TOF-MS to detect proteomic differences before and after infection.The chip detected a series of differentially expressed protein peaks.Interestingly,both peaks at 16 912 Da and 17 581 Da corresponded precisely with the molecular mass of ISG 15,which may participate in antiviral activity during the process of infection.Thus,the results we obtained can serve as a basis to study the pathogenesis of HSV-1 and the interaction between the virus and its host.In addition,they can help in the discovery of new therapeutic targets for treatment of HSV-1 infection.