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Braz. j. infect. dis ; 17(4): 410-417, July-Aug. 2013. ilus, tab
Artículo en Inglés | LILACS | ID: lil-683127

RESUMEN

BACKGROUND: Mitogen-activated protein kinase (MAPK) signaling pathway plays an important role in response to viral infection. The aim of this study was to explore the function and mechanism of MAPK signaling pathway in enterovirus 71 (EV71) infection of human rhabdomyosarcoma (RD) cells. METHODS: Apoptosis of RD cells was observed using annexin V-FITC/PI binding assay under a fluorescence microscope. Cellular RNA was extracted and transcribed to cDNA. The expressions of 56 genes of MAPK signaling pathway in EV71-infected RD cells at 8 h and 20 h after infection were analyzed by PCR array. The levels of IL-2, IL-4, IL-10, and TNF-α in the supernatant of RD cells infected with EV71 at different time points were measured by ELISA. RESULTS: The viability of RD cells decreased obviously within 48 h after EV71 infection. Compared with the control group, EV71 infection resulted in the significantly enhanced releases of IL-2, IL-4, IL-10 and TNF-α from infected RD cells (p < 0.05). At 8 h after infection, the expressions of c-Jun, c-Fos, IFN-i, MEKK1, MLK3 and NIK genes in EV71-infected RD cells were up-regulated by 2.08-6.12-fold, whereas other 19 genes (e.g. AKT1, AKT2, E2F1, IKK and NF-κB1) exhibited down-regulation. However, at 20 h after infection, those MAPK signaling molecules including MEKK1, ASK1, MLK2, MLK3, NIK, MEK1, MEK2, MEK4, MEK7, ERK1, JNK1 and JNK2 were up-regulated. In addition, the expressions of AKT2, ELK1, c-Jun, c-Fos, NF-κB p65, PI3K and STAT1 were also increased. CONCLUSION: EV71 infection induces the differential gene expressions of MAPK signaling pathway such as ERK, JNK and PI3K/AKT in RD cells, which may be associated with the secretions of inflammatory cytokines and host cell apoptosis.


Asunto(s)
Humanos , Enterovirus Humano A/genética , Proteínas Quinasas Activadas por Mitógenos/genética , Rabdomiosarcoma/virología , Citocinas/genética , Ensayo de Inmunoadsorción Enzimática , Enterovirus Humano A/enzimología , Enterovirus Humano A/fisiología , Regulación Neoplásica de la Expresión Génica , Proteínas Quinasas Activadas por Mitógenos/fisiología , Reacción en Cadena de la Polimerasa , Rabdomiosarcoma/enzimología , Rabdomiosarcoma/genética , Transducción de Señal/genética , Transducción de Señal/fisiología , Factores de Tiempo , Células Tumorales Cultivadas , Regulación hacia Arriba , Replicación Viral
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