Objective:To investigate the vasculogenic mimicry formation induced by hypoxia in Ⅱ-Ⅲ human glioma cell and the effect of alphastatin peptide suppressing the hypoxia-induced vasculogenic mimicry formation and the mechanism.Methods:MTT

Transwell and three-dimentional culture were used to detect the proliferation

migration and tubule formation of SHG44.The expression of vascular endothelial growth factor-α（VEGF-α）

erythropoietin-producing hepatocellular carcinoma-A2 （EphA2） and matrix metalloproteinases 2 （MMP2） was detected by RT-PCR and Western blotting analysis.Results:The OD 490 in hypoxia group was 0.60±0.06 and in control group was 0.46±0.05.The number of cell migration was 178.71±18.81 in hypoxia group and 85.86±17.92 in control group.The tubule formation was 56.80±12.21 in hypoxia group and 4.20±2.62 in control group.The proliferation

migration and tubule formation in hypoxia group were significantly higher than that in control group.The expression of VEGF-α

EphA2 and MMP2 was upregulated in hypoxia.When various concentrations of alphastatin （100

1 000

10 000 nmol/L） were added to hypoxia group

the numbers of cell migration were 142.57±12.12

92.71±17.68

30.00±7.72 and the tubule formation were 47.71±10.58

18.86±8.40

8.43±5.62.The cell migration and tubule formation were significantly suppressed by alphastatin in a dose-dependent manner.In alphastatin group

the phosphorylation of EphA2 protein （P=0.037

F=4.629） and activation of MMP2 protein （P=0.005

F=9.331） were significantly suppressed but there was no change in VEGF-α protein.Conclusion:Ⅱ-Ⅲ human glioma cell is able to form vasculogenic mimicry induced by hypoxia and alphastatin peptide can suppress the hypoxia-induced vasculogenic mimicry.VEGF-α induced EphA2 phospharilation and MMP2 activation maybe the key pathway to form vasculogenic mimicry.