RESUMO
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.
Assuntos
Humanos , Antibacterianos , Farmacologia , Apoptose , Genética , Doxiciclina , Farmacologia , Regulação da Expressão Gênica , Genética , Células HeLa , Proteínas Imediatamente Precoces , Genética , Metabolismo , Plasmídeos , Genética , Transativadores , Genética , Metabolismo , Proteína Supressora de Tumor p53 , MetabolismoRESUMO
The objective of present study is to investigate the effect of human cytomegalovirus (HCMV) infection on human hippocampus neural stem cells NSCs differentiation in vitro, Fetal hippocampus tissue was dissociated mechanically and then cultured in proliferation medium with EGF and bFGF. Immunofluorescence method was used to detect the expression of NSCs marker-Nestin within these cells. Cultured in 10% FBS, NSCs began to differentiate. On the onset of the differentiation, HCMV AD169 (MOI=5) was added into the differentiation medium. After 7 days differentiation, the effect of HCMV infection on NSCs differentiation was observed by detecting the rate of nestin, GFAP and HCMV immediate-early (IE) positive cells with confocal microscopy and immunofluorescence method. The resucts showed most of the cells (passage 4-6 ) were Nestin positive and could differentiate into NSE-positive neurons and GFAP-positive astrocytes. On day 7 postinfection, 86% +/- 12% of infected cells were IE positive. The percentage of Nestin-positive cells was 50% +/- 19% and 93% +/- 10% (t= 6.03, P<0.01)and those of GFAP-positive cells was 81% +/- 11% and 55 +/- 17% (t=3.77, P<0.01) in uninfected and infected cells respectively. These findings indicated that NSCs were HCMV permissive cell and HCMV AD 169 infection suppressed the differentiation of Hippocampus-genetic human neural stem cells into astrocytes.
Assuntos
Humanos , Astrócitos , Biologia Celular , Diferenciação Celular , Células Cultivadas , Citomegalovirus , Fisiologia , Fator de Crescimento Epidérmico , Farmacologia , Fator 2 de Crescimento de Fibroblastos , Farmacologia , Hipocampo , Biologia Celular , Proteínas de Filamentos Intermediários , Metabolismo , Microscopia de Fluorescência , Células-Tronco Multipotentes , Biologia Celular , Metabolismo , Virologia , Proteínas do Tecido Nervoso , Metabolismo , Nestina , Neurônios , Biologia CelularRESUMO
In view of the predominance problems in present experimental teaching of medical microbiology, we reformed the experimental contents and experimental teaching methods,to establish an entire experimental curriculum teaching system to be suitable for seven-year system and long educational system students,which manifests the creative teaching idea.