Objective To investigate the effect of isoliquiritigenin on the proliferation and differentiation of SHG44 human brain glioma stem cells and related mechanisms.Methods SHG44 cells were divided into dimethyl sulfoxide (DMSO) control group, isoliquiritigenin (10-160 μmol/L) induction group, N-[N-(3,5-difluorophenacetyl)-1-alanyl]-S-phenylglycine t-butyl ester (DAPT, 2.0 μmol/L) antagonist group, and isoliquiritigenin (10-160 μmol/L) + antagonist DAPT (2.0 μmol/L) group. CCK-8, immunofluorescent staining, Western blot, and real-time PCR were used to measure cell inhibition rate and expression of differentiation proteins and Notch1 pathway genes.Results Within 12-48 hours, the cell inhibition rate of isoliquiritigenin decreased with the increasing concentration (P<0.05), and the number of stem cells decreased with the increase in differentiated cells; after 72 hours, cell inhibition rate increased with the increasing concentration (P<0.05), and there were reductions in differentiated cells and stem cells. The drugs were added every other day, and on the seventh day, there were significant reductions in the number and diameter of glioma stem cells (compared with the control group, P<0.05). After 72 hours of isoliquiritigenin treatment, the expression of Nestin protein was gradually downregulated with the increasing concentration of isoliquiritigenin (compared with the control group, P<0.05); compared with the control group, the 10, 40, and 160 μmol/L groups had a significant increase in the expression of GFAP protein (P<0.05), and the 40 μmol/L group had significantly higher expression of GFAP protein than the other groups (P<0.05); compared with the control group, the 10, 40, and 160 μmol/L groups had a significant increase in the expression of β-Tubulin III protein (P<0.05), and the 10 μmol/L group had significantly higher expression of β-Tubulin III protein than the other groups (P<0.05). After the treatment with Notch1 pathway antagonist, the isoliquiritigenin groups and antagonist group had significant reductions in the expression of Notch1, RBP-JK, and Hes1 genes compared with the control group (P<0.05); compared with the isoliquiritigenin group, the isoliquiritigenin+DAPT group had significant reductions in the expression of Notch1, RBP-JK, and Hes1 genes (P<0.05); compared with the antagonist group, the isoliquiritigenin+DAPT group had significant reductions in the expression of Notch1, RBP-JK, and Hes1 genes (P<0.05).Conclusions Isoliquiritigenin can induce the differentiation of SHG44 human glioma stem cells into astrocytes and neurons and inhibit the proliferation of glioma stem cells, possibly by downregulating the expression of Notch1, RBP-JK, and Hes1 in the Notch1 signaling pathway.