Foxg1 Inhibits Angioensis In Hepatocellular Carcinoma In Vitro

Objictive To investigate the effect of forkhead box G1(Foxg1)on the angiogenesis of human umbilical vein endothelial cells(HUVECs)by interfering and overexpressing Foxg1 in hepattomacells.It directly reflects the effect of Foxg1 on the angiogenic ability of hepatoma cells and explore its underlying mechanisms.Methods1.Western blot detection of Foxg1 expression in hepatoma cell lines.2.The conditioned medium of each liver cancer cell was extracted,and Transwell assay was used to detect the effect of Foxg1 on the migration of HUVEC cells.3.The hepatocellular carcinoma cell lines Hep3 B and Huh7 were used in this experiment,and after corresponding transfection,the cells were divided into 4 groups,that is,Hep3 B shfoxg1 group(Foxg1silence),Hep3 B control group,Huh7 expfoxg1 group(Foxg1overexpression)and Huh7 control group.4.After the HUVECs were treated with the conditioned mediums(CM) from the above 4groups of cells,transwell assay and tube formation assay were used to observe the effect of CM on migration and tube formation of HUVECs.5.The mRNA levels of vascular endothelial growth factor A(VEGFA) in each group was measured by real-time fluorescence quantitative PCR(RT-PCR),and the protein levels was detected by Western blot.The enzyme-linked immunosorbent assay was used to measure the conecentration of VEGFA in the CM.Finally,Western the AKT phosphorylation level in each group of liver cancer cells was detected by Western blot.Results1.In the 7 hepatoma cell lines,Hep3 B had the highest Foxg1 expression, and Huh7 had the lowest Foxg1 expression.2.In 7 liver cancer cell lines,Hep3 B CM had the weakest migration ability to HUVEC cells,and Huh7 CM had the strongest migration ability to HUVEC cells.3.Transwell and luminal formation assays showed that under the action of CM in Hup3 B interference group and Huh7 control group,the migration ability and luminal formation of HUVEC cells were significantly enhanced,while in action of CM in Hep3 B control group and Huh7 overexpression group,The migration ability and lumen formation of HUVEC cells were significantly decreased(P<0.01).4.RT-PCR experiments showed that VEGFA levels were up-regulated in the Hep3 B interference group compared with the Hep3 B control group,and the levels were down-regulated in the Huh7 overexpression group compared with the Huh7 control group;ELISA experiments showed that VEGFA levels were up-regulated in the Hep3 B interference group CM compared with the Hep3 B control group CM,and the levels were down-regulated in the Huh7 overexpression group CM compared with the Huh7 control group CM(P<0.01).Western blot experiments showed that VEGFA and AKT1 phosphorylation levels were up-regulated in the Hep3 B interference group compared with the Hep3B control group,and the levels were down-regulated in the Huh7 overexpression group compared with the Huh7 control group.Conclusion Foxg1 inhibits the angiogenesis of hepatocellular carcinoma,which maybe through suppressing the PI3K/AKT pathway and down-regulating the expression of VEGFA.