Abstract: In order to screen for differential proteins involved in tumor migration in glioma exosomes and to find targets that may inhibit tumor cell migration, the culture supernatant discrepancies of A172 and U251 glioma cell lines with different migration capabilities was explored. The results showed that A172-conditioned medium had a stronger migration-promoting effect, while exosomes were the main components to promote tumor migration. Subsequently, the proteomic analysis of exosomes using tandem mass tag (TMT) quantitative mass spectrometry showed that differential proteins (fold change≥2) were mainly enriched in three KEGG (kyoto encyclopedia of genes and genomes) signaling pathways: ribosomes, gap junctions, and ubiquitin-mediated protein degradation. A total of 50 exosome proteins with significant expression differences were obtained, which could be used as exosome detection markers for gliomas and potential targets for inhibiting tumor cell migration.