Objective: To observe the inhibitory effect of survivin specific RNA interference (RNAi) vectors on the survivin mRNA and protein expression of pancreatic cancer PANC-1 cells. Methods: The plasmid vectors encoding U6 promoter-driven siRNA against survivin and the negative control plasmid were transfected into the pancreatic carcinoma PANC-1 cells, respectively. The mRNA and protein expression of survivin were detected by RT-PCR and Western blot, respectively. Results: In the PANC-1 cells transfected with RNAi vectors against survivin, the 24 h and 48 h inhibitory ratios on survivin mRNA expression were (52.67± 3.51)% and (75.33±3.06)%, and those on protein expression were (58.00±3.61)% and (76.67±4.73)%, respectively. All the differences had statistical significance compared with the negative control cells (all P<0.05). Conclusion: The RNAi vectors with U6 promoter can effectively inhibit the survivin expression in pancreatic carcinoma PANC-1 cells.