Title: Screening of genes differentially expressed in HepG2 cells transfected with SBPl expressing plasmid DNA using DNA microarray
Abstract: Objective To study the differences in gene expression in human hepatoblastoma cell line HepG2 cells transfected with SBP1_expressing plasmid DNA and further elucidate its potential molecular biological function. Methods Sequence specific primers were designed and synthesized and the SBP1 DNA fragment was amplified with polymerase chain reaction (PCR) technique. The expressive vector of pcDNA3.1(-)_SBP1 was constructed. The HepG2 cells were transfected with pcDNA3.1 (-) and pcDNA3.1(-)_SBP1 using FuGENE 6 transfection reagent respectively. The mRNA was isolated and reverse transcribed. The cDNAs were subjected for microarray screening with 1152 cDNA probes.Results The expressive vector had been constructed and confirmed by restriction enzyme digestion and DNA sequencing analysis. High quality mRNA and cDNA had been prepared and successful microarray screening was conducted. The scanning results indicated that among 1152 genes which were obtained from gene expression profile analysis,there were 18 differences in which 12 genes were up-regulated and 6 genes were down_regulated in SBP1_expressing HepG2 cells.Conclusion cDNA microarray technology was successfully used to screen the genes differentially expressed in SBP1_expressing HepG2 cells, which bring some new clues for studying the potential molecular mechanism of SBP1 protein.
Publication Year: 2005
Publication Date: 2005-01-01
Language: en
Type: article
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