Abstract: Objective To investigate the mechanism of E26 transformation-specific sequence 4 (ETV4) affecting the proliferation and migration of colon cancer cells through the nuclear factor-κB (NF-κB) signaling pathway. Methods The expression level of ETV4 in normal colon tissues and cancer tissues was analyzed by the user-friendly interactive cancer transcriptome data analysis resource (UALCAN) database. Reverse transcription quantitative polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression level of ETV4 in normal intestinal epithelial cells and colon cancer cell lines. After silencing ETV4 in SW480 cells, qRT-PCR and Western blot were performed to detect the expression of ETV4 to assess transfection efficiency; colony formation and Transwell assays were conducted to explore the effects of ETV4 silencing on the proliferation and migration of colon cancer cells; the Western blot was used to detect the effects of ETV4 silencing on the protein expression of protein 65 (p65) and phosphorylated protein 65 (p-p65) in the NF-κB pathway. Results The UALCAN database analysis revealed high expression of ETV4 in colon cancer tissues. The qRT-PCR and Western blot showed that ETV4 expression was significantly higher in the colon cancer cell lines SW480, Lovo, Caco-2, and SW620 than in normal intestinal epithelial cells HIEC-6, with the highest expression in SW480 cells (P<0.001). Colony formation and Transwell assay results indicated that silencing ETV4 significantly inhibited the proliferation and migration of colon cancer SW480 cells (P<0.001). Western blot results showed that silencing ETV4 significantly inhibited the expression of p-p65 protein in the cells (P<0.001). Conclusion Silencing ETV4 may inhibit the activation of the NF-κB signaling pathway, thereby inhibiting the proliferation and migration of colon cancer cells.