Construction of human SET gene RNA interference recombinant plasmid vector and its expression in L-02 liver cells

Objective To construct human patient SE transloction(SET) gene RNA interference recombinant plasmid vector and investigate SET protein's expression in the liver cells.The aim of this study was to lay a good basis for the further study of effect of SET protein on trichloroethylene-induced liver cells.Methods The 2 pairs of short hair-pin RNA oligonucleotides targeting human SET gene were designed and synthesized.After annealing,these double-strand DNA were cloned to eukaryotic expression plasmid Vector psiRNA-hH1 neo to form the shRNA recombinant plasmid vector,and they were confirmed to be correctly constructed by digestion and sequencing,and then were used to transfect into L-02 liver cells.48 h after the transfection,the liver cells were harvested,and the expression of SET protein in the liver cells after RNAi was detected by western blotting.Results The digestion and sequencing results confirmed that the recombinant plasmid vectors were correctly constructed,and Western Blotting results showed that transfection of recombinant plasmid psiRNA1 significantly down-regulated SET protein expression in L-02 liver cells,with the inhibition ratio of approximately 60%,P0.05,which showed significant difference from the normal control group.Conclusion The human SET gene RNA interference recombinant plasmid vector was successfully constructed,which inhibited SET protein expression effectively,indicating that it laid the groundwork for the study of investigating the role of SET protein in the process of trichloroethylene-induced liver cells.