Objective To study the role of ubiquitin ligase FBW7 in the sensitivity of glioma to temozolomide and its mechanism in glioma cells. Methods FBW7 overexpression lentivirus was constructed. Glioma cell line U251 was divided into 4 groups: the control group, temozolomide group, FBW7 overexpression group, and FBW7 overexpression + temozolomide group. Compared with the intervention on U251 cell lines, the differences of cell inhibitory rates in 4 different groups were analyzed by using contrast microscopy and methyl thiazolyl tetrazolium (MTT) colorimetric assay after 36 h and 72 h respectively. Flow cytometry (FC) was used to determine the cell cycle and apoptosis rate. Results The survival number of U251 cells in the three treatment groups was increased compared with the control group at both 36 h and 72 h. The inhibitory rates of temozolomide group, FBW7 overexpression group, and FBW7 overexpression +temozolomide group at 36 h were (17.6±0.8) ％, (10.4±0.6) ％, (18.6±0.6) ％ respectively compared with the control group (F=67.02, P<0.01); while at 72 h, the inhibitory rates of the three treatment groups were (25.1 ±0.4) ％, (16.7 ±0.7) ％, (29.0 ±0.9) ％ respectively compared with the control group (F= 74.61, P<0.001). Moreover, FBW7 overexpression + temozolomide group presented much greater inhibitory rate than temozolomide group (P<0.01). The G2/M arrest ratio and the cell apoptotic rate at 72 h in the three treatment groups were higher than those in the control group (F=41.63, P<0.001;F=42.30, P<0.01). The increased degree of G2/M arrest ratio and the cell apoptotic rate in FBW7 overexpression + temozolomide group were more obvious compared with temozolomide group (P<0.05, P<0.01). Conclusion FBW7 could enhance the sensitivity of glioma cells to temozolomide treatment, which is associated with G 2/M arrest and the increased apoptosis rate induced by FBW7.