Recombinant Plasmid PEGFP-AFP-hTNF-α Delivered By Nano-magnetic Fluids Killed Liver Cancer Cells (HepG2 Cells) In Vitro

Objective:1.To identify the possibility of using nano-magnetic fluids as gene vector for liver cancer gene therapy.2.To study the effect of AFP enhancer on tumor-specific expression of foreign gene in AFP-producing cancer cells like HepG2 cells.Methods:Construct recombinant plasmid pEGFP-AFP-hTNFαwhich was delivered by nano-magnetic fuilds into AFP positive HepG2 cells and AFP negative Hela cells.The fluorescence was detected in order to evaluate the transfection rate after transfecting 12hs.RT-PCR and Westernblot were used to detect expression of hTNFαgene at 48h after transfection.MTT assay was used to evaluate the effect of hTNFαon the proliferation of HepG2 cells.Flow cytometry was used to analysis the anpoptosis of HepG2 cells.Results:Nano-magnetic fluids delivered plasmid pEGFP-AFP-hTNFαinto HepG2 cells and hTNFαgene was successfully expressed.The transfection efficacy of nano-magnetic fluids was superior than Lipofectamine in HepG2 cells.RT-PCR and Westernblot demonstrated that hTNFαgene was expressed in HepG2 cells after being transfected with nano-magnetic fluids/pEGFP-AFP-hTNFα.MTT and flow cytometry showed hTNFαgene exerts cell-killing effect. Conclusions:1 Nano-magnetic fluids could successfully deliver pEGFP-AFP-hTNFαinto HepG2 cells and induce expression of hTNFαgene,it is promising gene vector for liver cancer gene therapy.2 AFP enhancer ean specifically enhance the expression of target gene within the cell with positive AFP.