Effect Of Isoliquiritigenin On Glioma Stell Cells Invasion And Migration

Objective:In vitro studies of isoliquiritigenin(ISL)on Glioma stem cells(GSC)influence the invasion and migration capacity,the effects of in vivo experimental study on ISL bearing glioma stem cells in nude mice.To provide the experimental basis for the new way about ISL in the treatment of glioma tumor mechanism.Methods:1.SHG44 glioma cells were cultured in DMEM culture medium containing 10%fetal bovine serum.SHG44 glioma cells were extracted and purified from DMEM culture medium containing10%fetal bovine serum using DMEM-F12 culture medium without fetal bovine serum.At the same time,the dry characteristics of glioma stem cells were detected by immunofluorescence.2.The migration and invasion of SHG44 glioma cells were observed after treated with 20?mol/L and 80?mol/L isoglycyrrhizin in DMSO-20?mol/L and 80?mol/L by cell scratch and Transwell invasion assay.The expression of matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase 9(MMP-9)were analyzed by western-blot.3.Transwell migration and invasion experiments were used to observe the effects of isoliberrrhizin on the migration and invasion of SHG44 glioma stem cells after treated with 20?mol/L and 80?mol/L of DMSO-20?mol/L and 80?mol/L.The lexpression of RNA in MMP-2and MMP-9 was detected by Real-time PCR for 48 h after treatment with 20?mol/L and 80?mol/L isoglycyrrhizin in SHG44 glioma cells.At the same time,the expression of MMP-2 and MMP-9 in human brain were analyzed by western-blot.4.The xenotransplantation model was established in nude mice with SHG44 glioma stem cells 0.1 ml(10~6ml).After the establishment of the model,intraperitoneal injection of BALB/C,low dose isoglycyrrhizin,high dose isoglycyrrhizin and fixed dose of temozolidomide were used to establish the xenotransplantation model.The relative tumor proliferation rate,tumor volume change and tumor inhibition rate were compared before and after administration.Results:1.SHG44 glioma stem cells were suspended in serum-free medium and grew as clonal spheres.The glioma stem cells were round under inverted microscope and spherical under scanning electron microscope.The long fusiform protuberances and small protuberances were also observed.The glioma stem cells labeled CD133 and Nestin were expressed positively.2.After treated with DMSO-20?mol/L and 80?mol/L ISL,the migration and invasion ability of SHG44 glioma cells decreased.3.The expression of MMP-2 and MMP-9 decreased in a dose-dependent manner.3.After treated with 20?mol/L and 80?mol/L ISL,the migration and invasion ability of SHG44 glioma stem cells decreased.ISL inhibited the gene expression of MMP-2,MMP-9 and the protein level of MMP-2 in a dose-dependent manner.4.BALB/C xenotransplantation model was injected intraperitoneally with low and high doses of ISL and fixed dose of TMZ 21 days later.The relative tumor proliferation rate in the model treated with low and high doses of ISL was less than 100.The difference between the two groups was statistically significant(P<0.05).Within 21 days after administration,DMSOs,low dose and high dose ISL and fixed dose TMZ groups were significantly different(P<0.05).The tumor volume of BALB/C nude mice model was gradually increasing,and there was no significant difference in tumor volume in each group within 9 days,but the tumor volume increment in the high dose ISL group was smaller than that in the control group after 12 days(P<0.05),and the tumor volume increment in the administration group was lower than that in the control group(P<0.05).The tumor volume increment of high dose ISL was lower than that of low dose ISL on the same day.Conclusions:1.ISL can inhibit the migration and invasion of glioma stem cells by affecting the expression of MMP-2 and MMP-9 gene and protein.2.ISL can inhibit the proliferation of SHG44 glioma stem cells in nude mice.