ABSTRACT
BACKGROUND: Human cytomegalovirus (HCMV) is a virus which has the potential to alter cellular gene expression through multiple mechanisms. OBJECTIVE: With the application of DNA microarrays, we could monitor the effects of pathogens on host-cell gene expression programmes in great depth and on a broad scale. METHODS: Changes in mRNA expression levels of human endothelial-like ECV304 cells following infection with human cytomegalovirus AD169 strain was analyzed by a microarray system comprising 21073 60-mer oligonucleotide probes which represent 18716 human genes or transcripts. RESULTS: The results from cDNA microarray showed that there were 559 differential expressed genes consisted of 471 upregulated genes and 88 down-regulated genes. Real-time qPCR was performed to validate the expression of 6 selected genes (RPS24, MGC8721, SLC27A3, MST4, TRAF2 and LRRC28), and the results of which were consistent with those from the microarray. Among 237 biology processes, 39 biology processes were found to be related significantly to HCMV-infection. The signal transduction is the most significant biological process with the lowest p value (p=0.005) among all biological process which involved in response to HCMV infection. CONCLUSION: Several of these gene products might play key roles in virus-induced pathogenesis. These findings may help to elucidate the pathogenic mechanisms of HCMV caused diseases.
Subject(s)
Cytomegalovirus Infections/genetics , Cytomegalovirus/genetics , Endothelial Cells/virology , Gene Expression Profiling , China , Cytomegalovirus Infections/pathology , Female , Humans , Male , Polymerase Chain Reaction , Tissue Array AnalysisABSTRACT
Objective To investigate the mechanisms for cytopathic effect(CPE) of wide-type rubella virus(RV)in ECV304 endothelial-like cells.Method ECV304 cells were cultured in MEM supplemented with 10% fetal calf serum,glutamine,sodium bicarbonate,penicillin and streptomycin at 37℃ with 5% CO-2 in a humidified incubator,and then grown in maintenance medium with 2% fetal calf serum to avoid overstimulating cell growth.Infected cells and control cells were harvested at day 2,4, and 6 after infection for following experiments.RT-PCR and indirect immunofluorescence were used to detect RV infection.Phase-contrast and electron microscopy were performed to analyze morphology changes in infected and uninfected groups.TUNEL assay was carried out to study wide-type RV-induced apoptosis.Result In RV-infected ECV304 cells,cytopathic effects were first observed approximately at 2-3 days post-infection.The majority of cells with CPE changes were of detachment from the monolayer,rounding of cells,cell shrinkage and membrane blebbing were seen.At 4-6 days post-infection,the detached cells increased clearly;the CPE regions on the monolayer were extended,and the infected cells in the CPE regions showed in spindly form with wide spacing.RT-PCR and indirect immunofluorescence results indicated RV infection in ECV304 endothelial-like cells.Using electron microscopy,RV particles and its cytopathic effects including the condensed chromatin,fragmented nuclei and clustering of mitochondria with dramatic changes around nuclear were further examined.TUNEL results showed that the difference of apoptosis index between the infected group and uninfected group was significant(P
