Objective To determine the expression of the chemokine receptor CXCR4 in glioma cells, and to analyze the mechanism of CXCR4 influencing U251 cell migration under the treatment with CXCL12 by establishing an invasion and migration model.Methods Indirect immunofluorescence assay was used to determine CXCR4 expression in U251 cells, and flow cytometry was used to determine CXCR4 expression in different pathological grades of gliomas. An under-agarose cell migration assay model was established to explore the U251 cell invasion ability under different concentrations of CXCL12.Blank group and experimental group were set up.The experimental group was divided into five groups with CXCL12 concentration.The chemotaxis coefficient was calculated.Results CXCR4 protein was expressed in U251 cells. The expression rates of CXCR4 in grade I, Ⅱ, Ⅲ, and IV gliomas were 21.36%±2.70%, 26.39%±4.27%, 52.59%±2.37%, and 56.23%±1.24%, respectively; there were significant differences between the four grades of gliomas in the positive rate of CXCR4 (P<0.05). The cell migration distance was 40.85±5.16 μm for the control group, and 49±2.26 μm, 105.6±3.82 μm, 165.3±3.89 μm, 245.4±5.94 μm, and 161.45±3.18 μm, respectively, for the experimental groups. The cells showed significant migration at a factor concentration of 200-500 ng/mL as compared with that in the negative group (P<0.05).Conclusions CXCR4 protein is expressed in U251 glioma cells. The CXCR4 positive rate is higher with increasing pathological grades of glioma. An under-agarose cell invasion assay model is successfully established in the glioma cells, which provides a new method for the research on tumor cell migration. CXCL12 shows an obvious chemotactic effect on glioma, and the tumor cells directionally migrate along the factor concentration gradient.