Fan ZHANG, Xianquan ZHANG, Cheng QIAN. Foxg1 inhibits angiogenesis in hepatocellular carcinoma in vitro[J]. Journal of Third Military Medical University, 2018, 40(6): 494-499.
DOI:
Fan ZHANG, Xianquan ZHANG, Cheng QIAN. Foxg1 inhibits angiogenesis in hepatocellular carcinoma in vitro[J]. Journal of Third Military Medical University, 2018, 40(6): 494-499. DOI: 10.16016/j.1000-5404.201711040.
Foxg1 inhibits angiogenesis in hepatocellular carcinoma in vitro
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
2
The hepatocellular carcinoma cell lines Hep3B and Huh7 were used in this experiment
and after corresponding transfection
the cells were divided into 4 groups
that is
Hep3B shFoxg1 group (Foxg1 silence)
Hep3B control group
Huh7 expFog1 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-linked immunosorbent assay (ELISA). Finally
Western blotting was used to detect the protein levels of AKT1 and p-AKT1 in the 4 groups of cells.
Results
2
The number of cell migration was significantly larger in the HUVECs exposure to CM from the Hep3B shFoxg1 group than that from Hep3B control group (
P
<0.01)
while the number was smaller in the HUVECs treated with Huh7 expFoxg1 group CM than with Huh7 control group CM(
P
<0.01). The CM from the Hep3B shFoxg1 group promoted tube formation in HUVECs when compared with the CM from the Hep3B control group (
P
<0.01)
and the CM of the Huh7 expFoxg1 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 Hep3B shFoxg1 group than the Hep3B control group (
P
<0.01)
and that in the Huh7 expFoxg1 group was down-regulated than the Huh7 control group (
P
<0.01).Western blot analysis showed that the Hep3B shFoxg1 group had enhanced phosphorylation of AKT1 than the Hep3B control group
while the level was opposite in the Huh7 expFoxg1 group than the Huh7 control group.
Conclusion
2
Foxg1 inhibits the angiogenesis of hepatocellular carcinoma
which may be through suppressing the PI3K/AKT pathway and down-regulating the expression of VEGFA.
关键词
Keywords
references
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