〔Abstract〕 Objective To investigate the effect of miR-495-3p targeting budding uninhibited by benzimidazoles(BUB1) on signal transducer and activator of transcription 3(STAT3) signaling pathway on biological behavior of esophageal cancer(EC) cells. Methods The differentially expressed genes in EC tissues and normal tissues were screened by the cDNA microarray technique. The differentially expressed genes were analyzed by bioinformatics methods. The target genes of miRNAs were predicted by the TargetScan database and verified by a dual luciferase gene reporter assay. KYSE150 cells were divided into blank control group, NC mimics group and miR-495-3p mimics group. The activity of KYSE150 cells were detected by the CCK-8 method. Cell cycle and apoptosis were measured by flow cytometry. The expression ofBUB1mRNA was measured by real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR). The levels of BUB1, STAT3, phosphor(p)-STAT3, cyclin dependent kinase 1(CCNB1), cyclin dependent kinase 1(CDK1), B-cell lymphoma-2(Bcl-2), cysteinyl aspartate-specific proteinase-3(Caspase-3) and cysteinyl aspartate-specific proteinase-3(Caspase-9) were measured by Western blot. The migration and invasion abilities of the cells were measured by wound-healing and Transwell invasion assays. Results Differentially expressed genes were involved in biological processes, signaling pathways and network construction, which were mainly related to cell cycle.BUB1is the key(Hub) gene, andBUB1is the target gene of miR-495-3p.In vitroexperiments showed that overexpression of miR-495-3p could significantly inhibit the migration and invasion of EC cells and induce apoptosis and G2/M phase arrest. After treatment with overexpression of miR-495-3p, the expression of Caspase-3 and Caspase-9 in EC cells increased significantly(P<0.01), while the expression of Bcl-2,BUB1, CCNB1, CDK1 and p-STAT3 decreased significantly(P<0.01). Moreover, the STAT3 signaling pathway might play an important role in this process. Conclusion miR-495-3p may influence the biological behavior of esophageal cancer cells by down-regulating BUB1-mediated STAT3 signaling pathway.