Abstract: Objective To explore the hsa-miR-133a expression in the cell lines of human esophageal squamous cell carcinoma, predict and analyze the target genes of hsa-miR-133a using bioinformatic method and to provide theoretical guidance for the further function study. Methods The relative expressions of hsa-miR-133a were detected by real-time quantitative PCR in human esophageal squamous cell carcinoma cell lines KYSE-150,Eca109 and human normal esophageal epithelial cell line Het-1; hsa-miR-133a target genes were predicted by TargetScan, the PicTar and miRanda, then combined with the confirmed gene database DIANALAB-TarBase5.0 and miRTarBase, the intersection of the first three groups of forecast results were obtained for functional annotation and pathway enrichment analysis. Results Compared with normal esophageal epithelial cells, the expression levels of hsa- miR-133a in human esophageal squamous cell carcinoma cell lines KYSE-150, Eca109 were significantly lower;hsa-miR-133a target genes were signifi cantly enriched in the AKT and p53 signal pathway closely related with tumor. Conclusion hsa-miR-133a target genes may be involved in the regulation of pathogenic of esophageal squamous cell carcinoma.