Effect Of Glioma Stem Cell-derived Exosomes On The Biological Activity Of Vascular Endothelial Cells

Objective To study the effects of exosome from glioma stem cells(GSCs)on the proliferation,migration and hypoxia/reoxygenation injury of human brain microvascular endothelial cells(HBMECs),and to explore the role of GSCs-exo in Whether biological information can be transmitted between glial stem cells and vascular endothelial cells can lay a foundation for further study of the mechanism of action of exosomes in angiogenesis of glioma.Methods(1)Human glioma U87 cell line-derived glioma stem cells(GSCs)were cultured and isolated,and the cultured tumor spheroid cells and their differentiated cells were identified by immunohistochemical staining.Exosome was extracted and identified by QIAGEN kit.(2)Human brain microvascular endothelial cells(HBMECs)were treated with different concentrations of GSCs-exo(20?g/ml,40?g/ml,60?g/ml),and CCK-8 was used to detect Effects of vascular endothelial cell proliferation and Transwell chamber assay on cell migration ability;(3)Human brain microvascular endothelial cells(HBMECs)were randomly divided into control;H/R;H/R+GSCs-Exo groups.After treatment,the CCK-8 method was used to detect the proliferation of three groups of cells.Transwell chamber was used to detect the migration of cells in each group.Flow cytometry was used to detect the apoptosis of each group.Results(1)The tumorspheres were suspended in the culture state.Immunohistochemical staining showed that Nestin and CD133 were specificallyexpressed in the cultured tumorspheres.The staining of the cells induced by the tumorspheres showed positive expression of GFAP and Neun.The membranous vesicle structure with circular or elliptical shape under transmission electron microscopy was about 30 nm in diameter.It was confirmed by Westen-bloting test that CD63 was expressed in the extracted GSCs-exo.(2)With the increase of GSCs-exo concentration,the proliferative capacity of vascular endothelial cells increased gradually,and the migration ability was also greatly improved.The difference was statistically significant(P<0.05).(3)Compared with the CON group,the vascular endothelial cell viability in the H/R group was greatly reduced,the migration ability was significantly decreased,and the apoptosis rate was significantly increased(P<0.05).Compared with the H/R group,GSCs-exo significantly increased vascular cell viability,promoted cell migration,and inhibited vascular endothelial cell apoptosis(P<0.05).Conclusions(1)Successfully cultured glioma stem cells(GSCs)from human glioma cell line U87,identified them from stem cell markers(Nestin and CD133)and induced differentiation experiments(GFAP and Neun).GSCs derived exosomes(GSCs-exo)were identified.(2)Glioma stem cell-derived exosomes can promote the proliferation and migration of vascular endothelial cells,and GSCs-exo may be a key factor in angiogenesis of glioma.(3)In the glioma hypoxia/reoxygenation microenvironment,glioma stem cell-derived exosomes have protective effects on vascular endothelial cells.