〔Abstract〕 Objective To investigate the distribution characteristics and role of GRK5-Moesin pathway in glioma. Methods Glioma cell lines with GRK5 up-regulated/down-regulated were prepared by lentivirus transfection, which were identified by Western blot and qRT-PCR. Effect of GRK5 changes on Moesin expression was detected by Western blot. Immunofluorescence analyzed subcellular localization and distribution characteristics of GRK5 and Moesin in glioma tissues. Proliferation, migration, invasion and apoptosis of U87 cells were detected by CCK-8 assay, Cell scratch assay, Transwell assay and flow cytometry, respectively. Results GRK5, Moesin and CD44 were co-located in glioblastoma cell membrane. GRK5-Moesin were enriched in glioma niches and glioma blood vessels, but the distribution of GRK5 and Moesin in each blood vessel was different. Targeting GRK5-Moesin could affect the biological activity of U87 glioma cells. Conclusion GRK5-Moesin can promote the malignant progression of glioma and is closely related to glioma stem cells.