Colorectal cancer (CRC) is a common tumor type, and liver metastasis reduces the long-term survival in CRC patients. Natural killer (NK) cells play an important role in anti-tumor immunity. The aim of this study was to investigate the mechanism of miR-214-5p on NK cells in CRC liver metastasis. We collected clinical samples of CRC liver metastasis and nonmetastatic tissues and purchased the human NK cell lines NK92 and liver metastatic CRC cells KM12L4 for research. RT-qPCR, Western blot, CCK-8, Transwell, and flow cytometry methods were used to evaluate the effect of miR-214-5p/USP27X/Bim pathway regulating NK cell activity on CRC liver metastasis. In addition, we also investigated the potential targets and regulatory mechanisms of the signaling pathway of miR-214-5p. In this study, we found that miR-214-5p was downregulated in CRC liver metastasis tissues. After transfection of miR-214-5p mimic, the activity of NK cells was significantly enhanced, and the proliferation and migration ability of CRC liver metastasis cells were inhibited, while inducing tumor cell apoptosis. Further research proved that USP27X is a potential target for miR-214-5p and upregulates Bim level through deubiquitination. In addition, miR-214-5p mimic reduced the level of USP27X and Bim, thereby enhancing the antitumor effect of NK cells. In conclusion, our research results show that miR-214-5p promotes the antitumor effect of NK cells by regulating the USP27X/Bim pathway, thereby inhibiting CRC liver metastasis. This finding reveals the important role of miR-214-5p in regulating the immune function of NK cells, and provides new ideas for developing new immunotherapy strategies.