Abstract: Objective To construct expression vectors of small RNA interference aimed at survivin gene and to explore effect of survivin siRNA gene on proliferation and invasive ability of SW480 cell line. Methods A survivin siRNA plasmid （pRNAT/sur-siRNA）was constructed using pRNAT U6.1/Neo vector. After pRNAT/sur-siRNA vectors were transduced into SW480 cell line 48 hours, the expression level of survivin mRNA and protein were measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis, the effect of pRNAT/sur-siRNA vectors on SW480 cell line proliferation and invasive ability were evaluated by MTT assay and cell invasion experiment, respectively. Results survivin siRNA expression plasmid （pRNAT/sur-siRNA）was constructed successfully and pRNAT/sur-siRNA down-regulated expression level of survivin gene protein and mRNA dramatically in SW480 cell,85%,80% respectively. SW480 cell containing pRNAT/sur-siRNA vectors (sur-siRNA/SW480) was selected by G418 400μg/L. MTT assay showed proliferation of sur-siRNA/SW480 cell was inhibited obviously (inhibition ratio was 37.4%), lower than proliferation of control group cell (P<0.01). Cell invasion experiment showed that cell numbers of sur-siRNA/SW480, pRNAT/SW480 and SW480 cell, that penetrated membrance, were 153±66, 505±65 and 578±98, penetration numbers of sur-siRNA/SW480 cell was reduced obviously, compare to control groups cell (P<0.01). Conclusion The siRNA aimed at survivin gene could reduce the expression level of survivin gene protein and mRNA, and inhibit proliferation and invasive ability of SW480 cell line. The sequence of RNA interference against survivin may be a validity method to treat colorectal cancer.