Objective: To construct short hairpin RNA (shRNA) eukaryotic expression vectors targeting Livin andSurvivin genes, and to explore the impact of co-transfection of Livin and Survivin shRNA expression vectors onthe biological behavior of HepG2 cells.
Methods: shRNA eukaryotic expression vectors pSD11-Livin and pSD11-Survivin were designed and constructed then transfected into HepG2 cells separately or in combination. mRNAand protein expression in transfected cells was assessed by quantitative fluorescence PCR and Western blotting,respectively. Cell proliferation was measured by MTT assay and cell apoptosis by TUNEL assay.
Results: TheLivin and Survivin shRNA eukaryotic expression vectors were successfully constructed and transfected intoHepG2 cells. The relative mRNA expression levels of Livin and Survivin in HepG2 cells co-transfected withpSD11-Livin and pSD11-Survivin were 0.12 ± 0.02 and 0.33 ±0.13, respectively, which was significantly lower thanlevels in cells transfected with either pSD11-Livin or pSD11-Survivin (P<0.05). The relative protein expressionlevels of Livin and Survivin in the co-transfected cells were also significantly decreased compared to singletransfection(P<0.05). The inhibition rate of cell growth in the co-transfection group was higher than that in thesingle-transfection groups at 48 h, 60 h, or 72 h after transfection (P<0.01). The apoptotic rate increased to thegreatest extent in the co-transfection group relative to any other group (P<0.05).
Conclusions: Co-transfectionwith pSD11-Livin and pSD11-Survivin was more efficient than transfection with either vector alone in reducingthe mRNA and protein expression of Livin and Survivin genes in HepG2 cells. Co-transfection also inhibited theproliferation of transfected cells more than the other groups, and induced cellular apoptosis more effectively.