To apply the small interfering RNAs targeting survivin to inhibit expression of endogenous survivin gene in human hepatocarcinoma cell SMMC-7721, the recombinant plasmid pshRNA-survivins were transfectted into SMMC-7721. The expression level of survivin was determined by Western blot and immunofluorescence staining and the transcription of survivin gene was detected by semi-quantitative RT-PCR. The introduction of plasmids pshRNA-survivin was showed to efficiently and specifically inhibit the expression of survivin according to results of Western blot and immunofluorescence staining, with inhibitory rates at 62%～78％peaking 72 h. Semi-quantitative RT-PCR showed that mRNA transcription of survivin gene was reduced by nearly 57%～64%. On the contrast, the control plasmid did exhibit no inhibitory effect on the protein expression and mRNA transcription of survivin gene. The results demonstrate that the small interfering RNA targeting survivin gene shows dramastic inhibition on protein expression and RNA transcription.