Construction of siRNA Eukaryotic Expression Plasmids Targeting Human CD147 Gene and Their Effects on CD147 Gene Expression in HeLa Cells

Objective To construct the siRNA eukaryotic expression plasmids targeting human CD147 gene and observe their effects on CD147 gene expression in HeLa cells.Methods The specific oligonucleotides of siRNA targeting CD147 gene were designed and synthesized according to the CD147 gene sequence reported in GenBank and directionally cloned into plasmid pGenesil-1 with enhanced green fluorescence protein(EGFP)and Kan genes.The constructed recombinant plasmids were identified by restriction analysis and sequencing then transfected to HeLa cells in mediation of liposome, and their effects on mRNA transcription and protein expression of CD147 were observed.Results Restriction analysis and sequencing proved that three siRNA eukaryotic expression plasmids and their negative control plasmids were constructed correctly.The inhibitory rates of three siRNA eukaryotic expression plasmids on CD147 mRNA transcription in HeLa cells were 32.5%, 66.8% and 59.1%, and those to CD147 protein expression were 28.3%, 63.5% and 56.2%, respectively.Conclusion The siRNA eukaryotic expression plasmids targeting human CD147 gene were successfully constructed and showed significantly inhibitory effect on CD147 gene expression in HeLa cells, which laid a foundation of further study on function of CD147 gene.