Inhibition of GnT-V gene expression in BGC823 cell line by using Lentivirus-mediated shRNA

Objective To construct the shRNA expression vector which targeting human beta-1,6-N-acetylgluco -saminyltransferase(GnT-V) gene and to detect its silencing effects in human gastric cancer cell line BGC823.Methods Three pairs of target segments were synthesized and cloned into Lentivirus-mediated shRNA respectively and the vectors were identified by enzyme digestion analysis and DNA sequencing.Then the expression vectors were transfected into BGC823 cell line.The expressions of GnT-V gene at mRNA and protein levels were detected by Real-Time PCR and Western blotting.Results Enzyme digestion analysis and DNA sequencing showed that the target segments were cloned into pLVTHM lentiviral vector respectively.Real-Time PCR detected that the expression rate of GnT-V gene of BGC823 cells in RNA interference group had dropped 88.04%as compared with that of the control.Western blotting also showed that expression of GnT-V protein was decreased in RNA interference group.Conclusion The shRNA expression vector effectively inhibits the expression of GnT-V gene at mRNA and protein levels in BGC823 cell line.