The impact of miR-1247-5p/Smad2 axis on gastric cancer cell migration,invasion and epithelial-to-mesenchymal transition

WANG Jian; CHENG Xianyong; YU Ning

doi:10.11958/20240634

Abstract：Objective To explore the expression of miR-1247-5p in gastric cancer and its effect on migration and invasion of gastric cancer cells.Methods The differentially expressed miRNAs in adjacent non-cancerous tissue versus gastric cancer tissue of TCGA database were analyzed.The prognostic value of miR-1247-5p in malignant tumor was analyzed by receiver operating characteristic(ROC)curve.RT-qPCR assay was used to detect expression levels of miR-1247-5p of gastric cancer tissue and adjacent non-cancerous tissue collected in 10 cases from our hospital.Protein-Protein Interaction(PPI)analysis was used to identify downstream target genes of miR-1247-5p.Dual-luciferase reporter assay was used to observe the binding of miR-1247 to Smad2.Transwell assay was used to investigate the effect of miR-1247-5p overexpression on migration and invasion abilities of gastric cancer cells.RT-qPCR and Western blot experiments were used to examine the impact of miR-1247-5p on expression levels of Smad2 and epithelial-mesenchymal transition(EMT)marker molecules.Results The expression levels of miR-1247-5p were lower in gastric cancer tissue collected from both TCGA database and our hospital than those in adjacent normal tissue.ROC curve analysis indicated that the expression level of miR-1247-5p was an independent prognostic indicator for most tumor types.Overexpression of miR-1247-5p significantly inhibited the migration and invasion abilities of gastric cancer cells.Bioinformatics analysis suggested that the downstream target genes of miR-1247-5p were involved in EMT and other pathways closely related to tumor metastasis.PPI analysis revealed that Smad2 was a Hub gene in the EMT signaling pathway.Dual-luciferase reporter assay demonstrated the existence of binding sites between miR-1247-5p and Smad2.RT-qPCR and Western blot experiments confirmed that miR-1247-5p can inhibit the expression of Smad2 and the process of EMT.Conclusion miR-1247-5p is lowly expressed in gastric cancer tissue,and overexpression of miR-1247-5p can inhibit EMT signaling pathway and reduce the invasive metastatic potential of gastric cancer cells.