Effect of Notch1 pathway activation on migration and invasion of glioma stem cells

Abstract

Objective To investigate the effect of Notch1 pathway activation on migration and invasion of glioma stem cells(GSCs)and its potential molecular mechanism. Methods U87 cells were cultured in the stem cells medium; CD133-positive glioma cells were isolated by magnetic activated cell sorting method. GSCs were identified by immunofluorescence to detect CD133 and nestin. The recombinant lentivirus aimed at Notch1 silencing was prepared. GSCs derived from U87 were employed and assigned into three groups: shRNA-Notch1 group, shRNA-negative control group and control group(without any treatment). Real-time PCR was used to detect the Notch1 expression; Western blotting was used to detect the expressions of Notch1, Hes1, Akt, phosphorylated Akt(pAkt), mTOR and phosphorylated mTOR(pmTOR); the abilities of migration and invasion of GSCs were observed by Transwell cell migration and invasion assay. Results Immunofluorescence analysis showed that the GSCs tumor spheres could express neural stem cell marker CD133 and nestin. The mRNA and protein expressions of Notch1, and protein expressions of pAkt, mTOR, pmTOR in the shRNA-Notch1 group were significantly lower than those in the shRNA-negative control group and control group(P 0.05). Transwell assay showed that the migration and invasion abilities in the shRNA-Notch1 group were significantly reduced as compared with those in the shRNA-negative control group and control group(P<0.05). Conclusion The suppression of Notch1 pathway activity may regulate the migration and invasion of GSCs through PI3K pathway. Key words: Glioma stem cell; Notch1; Migration; Invasion