Foxg1 inhibits angiogenesis in hepatocellular carcinoma in vitro
ZHANG Fan;ZHANG Xianquan;QIAN Cheng;Department of Oncology,the Second Affiliated Hospital of Chongqing Medical University;Biotherapy Center,First Affiliated Hospital,Army Medical University (Third Military Medical University);
Objective 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 hepatoma cells,and explore its underlying mechanisms. Methods 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 sh Foxg1 group( Foxg1 silence), Hep3 B control group, Huh7 exp Fog1 group( Foxg1 overexpression) and Huh7 control group. After the HUVECs were treated with the conditioned mediums( CM) from the above 4 groups of cells,transwell assay and tube formation assay were used to observe the effect of CM on migration and tube formation of HUVECs. The mRNA and protein levels of vascular endothelial growth factor A( VEGFA) in each group were measured by real-time fluorescence quantitative PCR( RT-PCR) and Western blotting. The concentration of VEGFA in the CM was detected by enzyme-linkedimmunosorbent assay( ELISA). Finally,Western blotting was used to detect the protein levels of AKT1 and p-AKT1 in the 4 groups of cells. Results The number of cell migration was significantly larger in the HUVECs exposure to CM from the Hep3 B sh Foxg1 group than that from Hep3 B control group( P 0. 01),while the number was smaller in the HUVECs treated with Huh7 exp Foxg1 group CM than with Huh7 control group CM( P 0. 01). The CM from the Hep3 B sh Foxg1 group promoted tube formation in HUVECs when compared with the CM from the Hep3 B control group( P 0. 01),and the CM of the Huh7 exp Foxg1 group induced less formation than the CM from the Huh7 control group( P 0. 01). RT-PCR and Western blotting showed that the VEGFA level was higher in the Hep3 B sh Foxg1 group than the Hep3 B control group( P 0. 01),and that in the Huh7 exp Foxg1 group was down-regulated than the Huh7 control group( P 0. 01).Western blot analysis showed that the Hep3 B sh Foxg1 group had enhanced phosphorylation of AKT1 than the Hep3 B control group,while the level was opposite in the Huh7 exp Foxg1 group than the Huh7 control group.Conclusion Foxg1 inhibits the angiogenesis of hepatocellular carcinoma,which may be through suppressing the PI3 K/AKT pathway and down-regulating the expression of VEGFA.
【CateGory Index】： R735.7
【CateGory Index】： R735.7