Objective Bioinformatics was applied to screen differentially expressed microRNAs (miRNAs) in esophageal squamous cell carcinoma(ESCC) and predict their target genes and analyze the biological functions.Methods Twenty,pairs of esophageal squamous cell carcinoma tissues and normal paired tissues adjacent to the carcinoma were collected from June 2021 to February 2022, Southwest Medical University Hospital. The miRNA expression profile microarray data of ESCC was downloaded from the GEO database and normalized using the R software limma package to screen for differentially expressed miRNAs. The survival analysis of differentially expressed miRNAs was performed using the online tool Kaplan-meier Plotter. The expression of differential miRNAs associated with survival was verified in ESCC tissues by qRT-PCR; targetScan and miRDB were used to predict target genes, and DAVID was used to annotate GO functions and KEGG pathway enrichment analysis of target genes. The PPI network was constructed by STRING and Cytoscape to identify core genes. The cytoscape was used to construct miRNA-mRNA network diagrams. UALCAN was used to evaluate the expression of core genes in esophageal cancer. Results Compared with normal tissues, hsa-miR-93, hsa-miR-130b, hsa-miR-18a, and hsa-miR-23b were expressed up-regulated in ESCC tissues; while hsa-miR-1, hsa-miR-133b, and hsa-miR-378a were expressed down-regulated in ESCC tissues. Three of these down-regulated miRNAs were significantly differentially expressed in ESCC tissues compared with normal tissues. The expressions of core genes HNRNPK, HNRNPU, DHX15, MATR3 and HNRNPA3 were significantly increased in esophageal cancer, and these core genes are closely related to hsa-miR-1. Conclusion The miRNAs are differentially expressed in ESCC tissues. The miRNAs may be involved in regulating relevant biological functions and pathways through target genes, thus affecting the occurrence and development of ESCC