Background and Aims Long noncoding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1) is an oncogenic lncRNA that promotes the progression of various cancers through the competing endogenous RNA (ceRNA) mechanism. Using the TargetScan database, we previously identified binding sites between NEAT1 and microRNA-124-3p (miR-124-3p), as well as between miR-124-3p and catenin beta-1 (CTNNB1). Therefore, this study was conducted to investigate the expression of NEAT1, miR-124-3p, and CTNNB1 in pancreatic cancer and their interactions affecting pancreatic cancer cell functions.Methods A dual-luciferase reporter assay was used to validate the relationships among NEAT1, miR-124-3p, and CTNNB1. The expression levels of NEAT1 and miR-124-3p, as well as CTNNB1 protein expression, were detected in pancreatic cancer tissues and adjacent normal tissues, as well as in pancreatic cancer PANC-1 cells and normal pancreatic epithelial H6C7 cells. PANC-1 cells were transfected with NEAT1 siRNA alone or co-transfected with a miR-124-3p inhibitor. After transfection, changes in PANC-1 cell biological functions, epithelial-mesenchymal transition related protein expression, and tumor growth ability in mice were assessed.Results The dual-luciferase reporter assay confirmed the targeting relationships between NEAT1 and miR-124-3p, as well as between miR-124-3p and CTNNB1. NEAT1 and CTNNB1 expression levels were significantly upregulated, while miR-124-3p expression was downregulated in pancreatic cancer tissues (vs. adjacent tissues) and in PANC-1 cells (vs. H6C7 cells) (all P<0.05). NEAT1 siRNA transfection led to decreased NEAT1 and CTNNB1 expression and increased miR-124-3p expression in PANC-1 cells. However, co-transfection with a miR-124-3p inhibitor suppressed the expression changes in miR-124-3p and CTNNB1 (all P<0.05). NEAT1 siRNA transfection significantly reduced PANC-1 cell proliferation, migration, and invasion, while promoting apoptosis. Additionally, E-cadherin protein expression was upregulated, whereas N-cadherin and vimentin protein expression were downregulated. Tumor growth in mice was also significantly inhibited (all P<0.05). These changes were attenuated upon co-transfection with the miR-124-3p inhibitor (all P<0.05).Conclusion NEAT1 may act as a ceRNA by competitively binding to miR-124-3p, thereby attenuating miR-124-3p-mediated inhibition of CTNNB1. This leads to CTNNB1 upregulation, ultimately promoting the malignant biological behavior of pancreatic cancer cells.