Objective To investigate the effect of small interfering RNA (siRNA)-mediated Survivin knock-down on proliferation of human pancreatic cancer cell line Patu8988 and their sensitivity to Gemcitabi.Methods The siRNA against Survivin was constructed and transfected into Patu8988cells with LipofectamineTM 2000.The expression of Survivin was detected by RT-PCR and Western blot.Flow cytometry was used to detect the cell cycle.Sensitivity to Gemcitabi after transfection were examined by MTT and clonogenic assay.Results In Patu8988 cells,the protein and mRNA levels of Survivin were decreased significantly after transfection,gene expression:control group:0.78 ± 0.03,blank control group:0.82±0.06,experimental group:0.52 ± 0.05 ; protein expression were as follows:control group:0.77 ± 0.21,blank control group:0.77 ± 0.26,experimental group:0.57 ± 0.03,and the difference was statistically significant ( P ＜ 0.05 ).And reduction of proliferation was related to an increase in the fraction of G0/G1 phase.The sensitivity of Patu8988 cells to Gemcitabi was increased significantly after transfection.Conclusions The Survivin special siRNA silenced Survivin,decreased Patu8988 cells proliferation and enhanced their sensitivity to Gemcitabi.