MicroRNA (miRNA) is a key regulator in the processes of disease development, however its regulatory mechanism in inflammatory reaction, immune response, especially in tumors is still unknown. In this study, we firstly predicted and analyzed the location, sequence conservation of hsa-miR-146a-5p aiming to get deep insight into the expression level in different tissues. Meanwhile, the target genes of hsa-miR-146a-5p were predicted by Targetscan7.1, miRDB, mirDIP and DIANA TOOLS. Additionally, protein-protein interaction (PPI) network was constructed through String 11.0. Gene ontology (GO) and Kyoto encyclopedia of genes and genome (KEGG) pathway enrichment analysis were performed on Omicshare platform and KOBAS databases. The results have indicated that the mature sequences of hsa-miR-146a-5p was highly conserved, with important biological functions. The 73 target genes intersect mainly involved transcriptional activity, structural molecular activity, catalytic activity, and activation of transcription factor active ligands. The significantly enriched biological processes included metabolism, signal transduction, positive or negative regulation, and they were related to Toll-like receptors pathway, NF-κB pathway, RIG-I like receptors and other signaling pathways. Our study was preliminarily conducted to elucidate that hsa-miR-146a-5p participates in tumorigenesis, immune response and inflammatory stimulation by regulating target genes, which would be a novel biomarker and therapeutic target.