Ovalitenin A, a chalcone compound isolated from the roots of Millettia brandisiana Kurz, has notable cytotoxic activity across various cancer types. Nevertheless, its therapeutic potential in cholangiocarcinoma (CCA), an aggressive malignancy arising from bile duct epithelial cells, remains underexplored. This study, therefore, seeks to evaluate the anti-metastatic properties of ovalitenin A and elucidate its molecular mechanisms in CCA cells. Cell viability and migratory capacity were evaluated using sulforhodamine B (SRB) and wound-healing assays, respectively, while western blot analysis was employed to assess the expression of key components in the NF-κB signaling pathway and its downstream targets. Results demonstrated that ovalitenin A dose-dependently inhibited CCA cell proliferation and significantly reduced cell migration. Furthermore, treatment with ovalitenin A upregulated the expression of IκBα and NF-κB proteins. Concurrently, the expression of vascular endothelial growth factor (VEGF), a critical NF-κB-regulated protein implicated in cancer metastasis, was significantly reduced upon ovalitenin A treatment. In conclusion, these findings provide compelling evidence for the potent anti-metastatic effect of ovalitenin A in CCA cells, highlighting its impact on inhibiting migration through modulation of the NF-κB signaling pathway.