Mechanism of Mir-217 in Suppressing Pancreatic Cancer Progression by Targeting ENO1

This research was designed to investigate the functional impact of miR-217 on proliferative activity and apoptotic processes in pancreatic carcinoma cells, and to explore its associated molecular mechanisms. Five matched pairs of pancreatic ductal adenocarcinoma specimens and corresponding adjacent non-neoplastic tissues were obtained. Quantitative real-time PCR (qRT-PCR) was utilized to assess the expression levels of miR-217. In the PANC-1 cellular model, gain-of-function and loss-of-function strategies were implemented through transfection with an miR-217 mimic or an antisense oligonucleotide (ASO), respectively. Plasmid-based overexpression and short hairpin RNA (shRNA)-mediated knockdown were utilized to modulate ENO1 expression. Cell proliferation was assessed using the MTT and colony formation assays. Apoptotic rates were quantified using flow cytometric analysis. Western immunoblotting was performed to determine protein expression of ENO1, AKT1, N-cadherin, E-cadherin, and Vimentin. The direct binding of miR-217 to the 3′-untranslated region (3′-UTR) of ENO1 mRNA was confirmed using a dual-luciferase reporter system. Functional rescue studies were conducted to evaluate the regulatory relationship between miR-217 and ENO1. miR-217 expression was markedly downregulated in pancreatic tumor tissues. Ectopic overexpression of miR-217 suppressed PANC-1 cell proliferation and induced apoptosis. At the mechanistic level, miR-217 upregulation led to reduced expression of ENO1, AKT1, N-cadherin, and Vimentin, accompanied by elevated E-cadherin levels. The dual-luciferase assay confirmed direct binding of miR-217 to the 3′-UTR of ENO1. Restoration of ENO1 expression counteracted the suppressive influence of miR-217 on proliferation and its promotive effect on apoptosis. miR-217 is underexpressed in pancreatic cancer and functions as a tumor suppressor by directly targeting ENO1. This regulation is associated with altered expression of AKT1 and epithelial–mesenchymal transition (EMT) markers, which may contribute to the inhibition of proliferation and promotion of apoptosis in pancreatic cancer cells.