Abstract: Objective To construct a series of eukaryotic expression vector which can transcript shRNAagainst human skp2 gene and investigate the inhibitory effect of siRNAon skp2 expression in pancreatic caner cell line Capan-2.Methods Three pairs of hairpin-like oligonucleotides specific for human skp2 gene were designed and synthesized.The annealed oligonucleotides designed contain 5'-BamH Iand 3'-EcoR Ⅰ overhangs.Then they could be ligated into the eukaryotic expression RNAi-Ready pSIREN-RetroQvector.The recombined plasmids were verified by digestion with restriction endonuclease.Results The shRNAsequences were successfully inserted into the eukaryotic expression vector pSIREN-RetroQ.The sequence-specific siRNAefficiently down regulated the SKP2 expression at both mRNAand protein levels.The inhibition rate was 64.16% at mRNA level and 60.26% at protein level,respectively.Conclusion Three siRNA expression vectors were successfully constructed.The siRNAplasmid expression vectors against SKP2 can inhibit the SKP2 expression in Capan-2 cells efficiently.RNAi-mediated gene silencing of SKP2 can be a novel modality in gene therapy of pancreatic cancer.

Key words: RNA interference, SKP2, pancreatic cancer