〔Abstract〕 Objective To investigate the effect and mechanism of Urolithin B(UB) on proliferation, migration, invasion, apoptosis and cell cycle of glioblastoma cells. Methods The effects of UB on the proliferation, migration and invasion of glioblastoma U251 cells were detected by CCK-8, clone formation assay, scratch healing assay, Transwell invasion assay respectively. The regulation effect of UB on cell cycle and apoptosis was detected by flow cytometry. Western blot was used to detect the effects of UB on the phosphorylation levels of downstream signaling pathway proteins ERK, AKT, p38 and JNK. Results Compared with the control group, at 24 h and 48 h, the absorbance value of U251 cells was decreased by UB(P<0.01) in a dose dependent manner. UB reduced the percentage of cell clone formation(P<0.01). The percentage of the scratch healing area was reduced by UB(P<0.05 orP<0.01). The percentage of the number of invaded cells was reduced by UB(P<0.01). UB could induce apoptosis(P<0.01) and cause the cells to stagnate in G2/M phase(P<0.01). UB could significantly inhibit the phosphorylation of ERK(P<0.05 orP<0.01) and AKT(P<0.05 orP<0.01). Conclusion UB can inhibit proliferation, migration, invasion, apoptosis and cell cycle of glioblastoma cells, and regulate the phosphorylation levels of ERK and AKT.