Characterization of differentially expressed human gene transcripts in uninfected versus dengue virus-infected ECV304 endothelial-like cells

Abstract

The complex changes in host cellular gene expression profiles at various stages of dengue virus infection were elucidated via differential display RT-PCR (DD-RT-PCR) and microarray screenings to analyze changes in mRNA expression patterns in the human endothelial-like ECV304 cell line. 78 distinct clones of differentially expressed ESTs were discovered in the DD-RT-PCR experiment while 148 differentially expressed genes were found in the microarray screen. Selected genes of various functions were validated via semiquantitative RT-PCR and real-time RT-PCR. The DD-RT-PCR data were generally congruent and in agreement with the semiquantitative RT-PCR and validated real-time RT-PCR data. However, the microarray data showed only 53% agreement with the validated real-time RT-PCR data thus highlighting the variability and requirement for validation of such high-throughput screens. In addition, CD9, CD63 and CD81 receptors were found to be highly expressed in ECV304 cells but antibody blocking experiments did not yield any statistically significant reduction in infectivity.