Background: Epithelial-mesenchymal transition (EMT) is a major actor modulating the metastasis of nasopharyngeal carcinoma (NPC). Increasing evidence indicates that microRNAs (miRs) are the important regulators of EMT program. However, the potential roles and underlying mechanisms of miR-379-5p in regulating EMT of NPC cells remain unclear. Methods: miR-379-5p expression levels in human NPC tissues and cell lines were detected via quantitative real-time PCR (qRT-PCR). Then, the correlations between miR-379-5p expression in NPC tissues and clinicopathologic features and patients' prognosis were analyzed. The effect of miR-379-5p on the expression of EMT markers in NPC cells was evaluated by Western blot and qRT-PCR. NPC cells' migration and invasion were evaluated in vitro by Transwell migration and invasion assays, respectively. The target of miR-379-5p was predicted with three publicly available databases and further validated with dualluciferase reporter assay, qRT-PCR, and Western blot. Results: The expression of miR-379-5p was significantly decreased in NPC tissues, and its low expression was significantly associated with multiple unfavorable clinicopathological factors and poor prognosis of NPC patients. Meanwhile, miR-379-5p was downregulated in NPC cell lines, and its exotic expression inhibited EMT to reduce the migration and invasion of NPC cells. Furthermore, Y-box binding protein 1 (YBX1) was identified and validated as a direct target of miR-379-5p, and restoring YBX1 expression could reverse the inhibitive effect of miR-379-5p on NPC cell EMT, migration and invasion. Conclusion: Taken together, our findings indicate that miR-379-5p inhibits the EMT of NPC cells to reduce their migration and invasion abilities by post-transcriptionally suppressing YBX1 expression, providing a novel potential treatment target for NPC patients.