Rectal cancer seriously threats to human health. Traditional chemotherapy drugs might kill rectal cancer cells while easy cause side effects and clinical complications. Therefore, it is necessary to explore possible new methods for rectal cancer treatment. Survivin is an important tumor-specific protein. Previous researches showed it may be closely related to nasopharyngeal carcinoma. Its role in rectal cancer remains unclear.Cultivate human rectal cancer HRC-9698 cells. Antisense microRNA targeting survivin and control miRNA were constructed and transfected to HRC-9698 cells. MTT assay was applied to detect cell growth. Flow cytometry was used to test cell apoptosis. Western blot was performed to detect Osteopontin expression level. Colony formation and transwell assay were used to test cell clone formation and invasion abilities.Antisense microRNA targeting survivin can inhibit HRC-9698 cell proliferation and induce its apoptosis. Antisense microRNA targeting survivin decreased osteopontin expression level. Overexpressed osteopontin inhibited antisense microRNA targeting survivin induced cell apoptosis. Antisense microRNA targeting survivin suppressed HRC-9698 cells' colony formation and invasion abilities.Antisense microRNA targeting survivin induced osteopontin participated HRC-9698 cell apoptosis. Antisense microRNA targeting survivin inhibited HRC-9698 cell colony formation and invasive abilities, indicating that survivin may play its anti-tumor effect through inducing cell apoptosis and inhibiting cell metastasis and invasion.