The objective of the study was to evaluate the expression of survivin, cell proliferation, and apoptosis in survivin-specific siRNA-transfected human gastric cancer cell line MGC-803. For this purpose, the target gene fragments were cloned into pSilencer3.1-Hl neo vector. Recombinant eukaryotic expression vector, pSilencer3.1-SVV was successfully constructed and then the recombinant vector was transfected into gastric cancer MGC-803 cells. The mRNA expression of survivin was determined by reverse-transcriptase polymerase chain reaction (RT-PCR). Survivin protein expression was detected by Western blot. Cell cycle distribution and apoptosis were determined by flow cytometry. Our data regarding RT-PCR and Western blot showed that pSilencer3.1-SVV vector could knockdown the expression of survivin mRNA and protein. In contrast with the control group, the apoptotic index of MGC-803 cells increased remarkably. Survivin-specific siRNA caused cells accumulation in the G2/M phase and the number of cells in the G0/G1 phase decreased after transfection. It was, therefore, concluded that the siRNA targeting survivin gene could inhibit the proliferation of gastric cancer cells and induce apoptosis. The use of survivin siRNA may provide a novel approach for gene therapy of gastric cancer.