Colorectal cancer is a common health-threatening tumor within the gastrointestinal tract. The aim of this study was to test the biological role of DUSP4 in colorectal cancer cells. In our study, DUSP4 overexpression-treated HCT116 cells and DUSP4 knockdown-treated SW480 cells were selected to perform study. Quantitative real-time PCR test (qRT-PCR) and western blot were used to detect DUSP4 abundance in clinical tissues and six cell lines, as well as ubiquitin-related Smad4 degradation. Western blot, migration and invasion. were used to assess the relationships between DUSP4 and Smad4. Higher DUSP4 expression of functional significance was observed in colorectal cancer tissues and cells. The results showed that both treatments could affect the proliferation, colony formation, migration, invasion of tumor cells, and the expression of epithelial mesenchymal transformation (EMT)-associated biomarkers. Moreover, in colorectal cancer cells, DUSP4 could promote the Smad4 degradation by regulating ubiquitin-related Smad4 degradation, and promote the cell proliferation, migration and invasion by regulating Smad4 degradation via Smad4 gene. Meanwhile, DUSP4 can directly deubiquitinate and stabilize Smad4 protein, hence further promote proliferation and metastasis of colorectal cancer cells.