Abstract: Objective To detect the role of VEGF₁₆₅ gene in vasculogenic mimicry formation of HepG2. Methods Two and three-dimensional culture was wed to observed the vasculogenic mimicry formation of HepG2.Eukaryotic expression plasmids pcDNA3.0-VEGF₁₆₅ and pcDNA3.0-VEGF₁₆₅b were constructed and transfected into HepG2 cell by Lipofectamine2000，then cell were cultured in the three-Dimensional matrix，the expression of VEGF₁₆₅ and VEGF₁₆₅b were detected by RT-PCR and Western blot. Results HepG2 cells could form vasculogenic mimicry in three-Dimensional matrix.The expression of VEGF₁₆₅ and VEGF₁₆₅b of HepG2 cells were up-regulation after transfected with their corresponding plasmids，but the vasculogenic mimicry formation of transfected HepG2 cells has no change. Conclusion HepG2 cells could form vasculogenic mimicry in three-Dimensional matrix，but the overexpression of VEGF₁₆₅ and VEGF₁₆₅b gene can not significantly influence the vasculogenic mimicry formation.