Objective: To investigate the effect of anterior gradient 2 (AGR2) on the migration and invasion of nasopharyngeal carcinoma (NPC) cells by regulating the transforming growth factor-β (TGF-β)/SMAD signaling pathway. Methods: Human normal nasopharyngeal epithelial cells (NP69) and NPC cells (5-8F, 6-10B, CNE-1) were cultured in vitro, and qRT-PCR was used to detect the expression of AGR2 mRNA in the cells. 5-8F cells were randomly assigned into Control group, sh-NC group, sh-AGR2 group, and sh-AGR2+SRI-011381 group (sh-AGR2+TGF-β signaling pathway activator SRI-011381). QRT-PCR and Western blot were applied to detect the transfection efficiency of AGR2. Cell proliferation was measured by CCK8 assay and colony formation assay. Cell migration was measured by scratch test. Transwell assay was used to measure cell invasion. Cell apoptosis was determined by flow cytometry. The protein expressions of PCNA, MMP-2, MMP-9, TGF-β1 and p-SMAD2/SMAD2 were detected by Western blot. Results: Compared with NP69 cells, the AGR2 mRNA expression was elevated in NPC cells. Compared with the Control group or sh-NC group, the OD450, clone number, scratch healing rate, invasion number, PCNA, MMP-2, MMP-9, TGF-β1, and p-SMAD2/SMAD2 expression levels in the sh-AGR2 group decreased, and the apoptosis rate was increased (P<0.05). Compared with the sh-AGR2 group, the OD450, clone number, scratch healing rate, invasion number, PCNA, MMP-2, MMP-9, TGF-β1, and p-SMAD2/SMAD2 expression levels in the sh-AGR2+SRI-011381 group increased, and the apoptosis rate was decreased (P<0.05). Conclusion: AGR2 is highly expressed in NPC cells, which can promote proliferation, migration, invasion, and inhibit cell apoptosis of NPC cells, and it may be related to the regulation of TGF-β/SMAD signaling pathway.