Colorectal cancer is a frequent malignancy, so its corresponding behaviors are essential to be understood deeply. This work was developed to explore the impacts of Livin on proliferation, migration, invasion, and epithelial-mesenchymal transition of HT-29 colorectal cancer cells. Colorectal cancer cells were extracted from pathological specimens collected from 80 colorectal cancer patients treated at our institution from June 2017 to June 2022. HT-29 human colorectal cancer cells were undertaken as the research materials. Real-time immunofluorescence polymerase chain reaction, Western blot, cell counting kit-8 experiments, cell scratch assays, and Transwell experiments were performed to assess the proliferation, migration, invasion, and epithelial-mesenchymal transition-related markers of the colorectal cancer cells. Livin expression in colorectal cancer cells increased greatly in contrast to that in healthy cells (p<0.05). Compared to normal HT-29, the overexpression of Livin promoted the proliferation, migration, and invasion abilities of colorectal cancer cells, exhibiting great differences (p<0.05). Moreover, levels of matrix metalloproteinase 2, vimentin, slug, and snail in colorectal cancer cells were sharply upregulated (p<0.05), while that of E-cadherin was remarkably downregulated (p<0.05) in contrast to the conditions in normal cells. The findings in this work revealed the crucial role of Livin in HT-29 colorectal cancer cells, suggesting its potential to be a target for colorectal cancer treatment.