Objective To investigate the relationship between Notch signaling pathway and ep-ithelial mesenchymal transition in glioma cells. Methods Immunomagnetic bead was used to isolate brainglioma stem cells from human glioma tissue. The glioma stem cells were cultured in vitro. Immunofluorescence assay was used to identify the cells. RNA interference retroviral vector of Notch-1 gene （pSiRNA-Notch-1）was constructed. The experiment contained three groups：blank control group （non-transfected plasmid），negativecontrol group （transfected with blank plasmid）and interference group （transfected with pSiRNA-Notch-1）. The proliferation and differentiation of three groups were observed. The expression of mRNA and protein of Notch-1 and Hes-1 was detected by RT-PCR and Western blot. Results The cell proliferation and cell differentiation absorbance（OD）of interference group after 7d were （1.332±0.614）and（1.203±0.783）respectively，which were significantly lower than that of blank control group and negative control group （P＜ 0.05）. The relative expression of mRNA and protein of Notch-1 and Hes-1 in interference group after 3 d proliferation was （0.189±0.021）and （0.301±0.121），（0.422±0.022）and（0.091±0.032），which was significantly lower than that in blank control group and negative control group （P＜ 0.05）. The relative expression of mRNA and protein of Notch-1 and Hes-1 in interference group after 3 d differentiation was （0.253±0.071）and（0.192±0.043），（0.178±0.022）and （0.101±0.012），which was significantly lower than that in blank control group and negative control group （P＜ 0.05）. Conclusions Notch signaling pathway plays an important role in proliferation and differentiation of glioma cells.Notch-1 and Hes-1 may be involved in the regulation of proliferation and differentiation，which needs further study.