Experimental Study Of Construction Of ShRNA Lentiviral Vector Targeting Human Nanog Gene And Trasfected Human Carcinoma Xenograft In Nude Mice

Objective:Build on the Nanog gene interference lentiviral vectorinvestigate its expression in human gastric cancer cell SGC-7901xenograftmodel, testing its effect on tumor growth in nude mice.Methods:The siRNA has been designed based on Nanog gene specificlentiviral vectors.Preparation of lentivirus carrying shRNA-Nanog,simultaneous construction of transplantation tumor model of human gastriccancer in nude mice.The lentivirus-shRNA-Nanog mice was used asexperimental group and the GFP infected mice and uninfected mice wereused as controls. The tumor volume and weight changes were measured.The Nanog protein expression was determined using Western blot analysis.The transinfected tumor cells apoptosis was analyzed using TUNELmethod.Results:The Nanog gene shRNA recombinant lentivirus wassuccessfully established. In vivo luminescence imaging showed that27 days after transfection, the total luminescence in the experimental groupwas significantly higher than the GFP infected and uninfected mice groups.The xenograft volume and weight in the experimental group wassignificantly lower than the GFP infected mice and uninfected mice groups.Western blot showed that the expression of Nanog protein was significantlylower than the control groups. TUNEL results indicated that the apoptoticrate in the experimental group was significantly greater than thecontrols.While there was no significant difference between the GFPinfected and uninfected mice groups.Conclusion:Lentivirus-shRNA-Nanog recombinant lentiviraltransfection of nude mice, the success of down regulated expression ofNanog protein, induced gastric cancer cell apoptosis, inhibit tumor growth,and lay the foundation for gene therapy of gastric cancer.