Objective: Survivin has attracted abundant interest in tumor research since it was discovered in 1997. But several studies indicated that the relationship between survivin expression and tumor behavior is still not fully understood. So our main aim was to observe the localization of survivin in colon cancer and its influence on the colon cancer biocharacteristics.Methods: We screened the expression and localization of survivin in SW480 by immunofluorescence and immunocytochemistry. Then, we constructed the recombinant adenovirus (Ad-survivin/shRNA), which contained the short hairpin RNA (shRNA)of survivin and transfected it into SW480 cells. We detected survivin gene expression after shRNA interference by RT-PCR and Western blot, and its influence on the proliferation, apoptosis and cell cycle was analyzed by MTT, colony-formation, and flow cytometry. Results: Survivin was expressed at a high level in SW480 cells and the sub-cellular localization was in the cytoplasm. Recombinant adenovirus could suppress survivin expression efficiency and inhibit proliferation, induce apoptosis,and affected the mitosis in vitro. Conclusion: Survivin plays an important role in controlling tumor growth by a variety of molecular regulatory mechanisms. Cytoplasmic survivin silence could induce apoptosis, effect the mitotic cycle and inhibit cell proliferation.