Mucin 1 (Muc1) is a tumor-associated glycoprotein and has an important role in cancer progression and metastasis. The aim of the study was to explore the effects and mechanism of Muc1 silencing on proliferation and tumor-forming capacity of colon cancer cell lines. In the present study, we first screened the colon cancer cell lines with high expression of Muc1 by western blot analysis. Then, the effective siRNA was selected and used for silencing endogenous Muc1. The results by MTT and vitro scratch assay showed that interference of Muc1 could effectively inhibit HCC2998 proliferation and migration (p<0.01). Also, colony-forming ability in Muc1-siRNA groups was significantly decreased compared with the control group (p<0.01). Cell cycle is detected by flow cytometry that suggested that Muc1-siRNA1 induced cell cycle arrest at S phase in HCC2998. Next, the expression and distribution of β-catenin in cytoplasm and the nucleus was detected by western blot and the results revealed that the expression of β-catenin was redistributed in Muc1-siRNA group. A higher β-catenin level was detected in cytoplasm, while a lower β-catenin level was located in nucleus, compared with controls (p<0.05). The tumorigenicity experiments showed that inhibition of Muc1 could significantly suppress the growth of HCC2998 in nude mice models (p<0.01). This study would effectively provide new clues for colon cancer therapy.