Construction Of Shrna Eukaryotic Expression Plasmid Targeting Human CD147 Gene And Their Effects On CD147 Gene Expression In HeLa Cell

Objective:To construct eukaryotic expression plasmids expressing short hairpin RNA(shRNA) sections targeting human CD147 gene and to observe their effects on CD147 gene expression in HeLa cell line.Methods:According to the sequence of human CD147 gene,the oligonucleotides of shRNA were designed and synthesized and directionally cloned into plasmid pGenesil-1 with enhancing green fluorescence protein(EGFP) gene and Kan gene.The recombinant plasmids were confirmed by enzyme digestion analysis and DNA sequencing.The recombinant plasmids were transfected into HeLa cell line by LipofectamineTM 2000,the effects on CD147 at mRNA and protein levels were observed.Results : Three shRNA expressing recombinants and the corresponding negative control plasmid were constructed and transfected into HeLa cell successfully.CD147 transcript was reduced by about 32.5%-66.8% ,the protein of CD147 was reduced by about 28.3%-63.5% in three transfectants respectively.Conclusion:The construction successfully of eukaryotic expression plasmids expressing shRNA sections targeting human CD147 and its noticeable inhibition on the expression of CD147 of HeLa cell line established a favorable foundation for further study on the function of CD147.