Glioblastoma multiforme (GBM) is a common primary tumor of the central nervous system. The change of cell stemness is one of the important characteristics of tumors including GBM, which is a cause and effect of tumor recurrence and metastasis. Upon binging to the receptor special CXCR4, CXCL12 is involved in the proliferation, invasion and metastasis of tumor cells. Here we report the impact on cancer stem cells (CSCs) in GBM. Bioinformatics software was used, based on TCGA, to analyze of the potential correlation between CXCL12-CXCR4 axis and stem-related genes (OCT4, SOX2, NANOG). Quantitative real time polymerase chain reaction (qRT-PCR), western blotting and sphere forming ability analyses were performed to access the effect of CXCL12-CXCR4 axis on GBM cell stemness in vitro and vivo.