Effects Of Isoliquiritigenin On Vasculogenic Mimicry Of SHG44 Human Glioma Cells In Hypoxic Environment

Objective:1.To investigate the effects of Vasculogenic mimicry and related biological behaviors(including proliferation and migration)of glioma cells in hypoxic environment induced by Cocl2.2.To investigate the effects of isoliquiritigenin on Vasculogenic mimicry of glioma cells in hypoxic environmentMethods1.SHG44 glioma cells were treated with Cocl2 at different concentrations(100,200,400uM)for 24 h,The vm of SHG44 cells was detected by threedimensional culture.The proliferation and migration of SHG44 cells were detected respectively by CCK-8and wound healing assay.The expression of VM related proteins such as EphA2,M MP-2 and VEGF were detected by Western blotting.2.SHG44 glioma cells were treated with low(20uM),medium(40uM),and high(80uM)concentrations of isoliquiritigenin for 24 hours under the hypoxic environment induced by Cocl2.The proliferation and migration of SHG44 cells were detected respec tively by CCK-8 and wound healing assay.The expression of VM related proteins suc h as EphA2,MMP-2 and VEGF were detected by Western blotting.Results1.The first part experiment1.1 Compared with the blank group,the number and length of vm were significantly increased when the concentration of Cocl2 was 100 uM and 200 uM.the cell proliferation activity and migration distance were increased,the difference was statistically significant(P<0.05).When the concentration of Cocl2 reached 400 uM,the number and length of vm decreased,and the cell proliferation and migration distance decreased.The difference was statistically significant(P<0.05).1.2 Compared with the blank control group,EphA2,MMP-2 and VEGF protein e xpressions were significantly increased when the concentration of Cocl2 was 100 uM a nd 200 uM,the difference was statistically significant(P<0.05).EphA2,MMP-2 and V EGF protein expressions were decreased when the concentration of Cocl2 reached 400 uM,the difference was statistically significant(P<0.05).2.The second part experiment2.1 Compared with the hypoxic blank control group,the number and length of VM were significantly decreased with the increase of isoliquiritigenin concentration,and the cell proliferation activity and migration distance were decreased,showing a dose-effect relationship,and the difference was statistically significant(P<0.05).2.2 The expression of EphA2 and MMP-2 proteins was inhibited by isoliquiritigen in in a dose-dependent manner,that is,the expression of EphA2 and MMP-2 proteins decreased with the increase of isoliquiritigenin concentration,and the difference was s tatistically significant(P < 0.05).In the hypoxic environment,there was no significant change in the expression of VEGF protein at the concentrations of 20 uM and 40uM(P > 0.05).When the concentration of isoliquiritigenin increased to 80 uM,the expres sion of VEGF protein decreased,and the difference was statistically significant(P < 0.05).Conclusions1.The first part experiment1.1Microhypoxic environment can induce the formation of VM in glioma,and at the same time,cell proliferation and migration ability were increased;When the degree of hypoxia was aggravated,VM formation and related biological behaviors(including proliferation and migration)can be inhibited.1.2Microhypoxia can up-regulate the expression of EphA2,MMP-2 and VEGFproteins.When the degree of hypoxia was aggravated,the expression of EphA2,MMP-2 and VEGF were inhibited.2.The second part experiment2.1 Isoliquiritigenin can inhibit vm formation and related biological behaviors(including proliferation and migration)Under the microhypoxia environment,and the mechanism may be by inhibiting the expression of EphA2,MMP-2 and VEGF proteins.