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Objective: The aim of this study was to explore the relationship between VEGF expression and vasculogenic mimicry of tumor in vitro. Methods: We observed the ability to form vasculogenic mimicry in several cell lines follow as LO2 cell, hepG2 cell, SMMC-7721 cell and A549 cell in three-dimensional cultures; we detected the expression of VEGF in the level of mRNA and protein by polymerase chain reaction (PCR) and western blot in those cell lines. Results: HepG2 cell and A549 cell had the ability to form vasculogenic mimicry in three-dimensional cell cultures, while LO2 cell and SMMC-7721 cell hadn't the ability. The rates of VEGF165/GAPDH were 0.212±0.011,0.208±0.013, 0.117±0.009 and 0.2±0.012 respec-tively, and the rates of VEGF121/GAPDH were 0.186±0.018, 0.192±0.014, 0.050±0.010, 0.196±0.017 in LO2 cell, hepG2 cell, SMMC-7721 cell and A549 cell, separately. The expression of VEGF gene in mRNA and protein levels in HepG2 cell, LO2 cell and A549 cell were significantly higher than that in SMMC-7721 cell (P < 0.05). Conclusion: The call lines which the expression of VEGF gene is low don't have the ability to form vasculogenic mimicry and the high expression of YEGF gene cenn' t form vasculogenic mimicry at all, while the expression of VEGF gene in the cell lines is high in the cell lines which can form vasculogenic mimicry. VEGF has impact on the formation vasculgenic mimicry. Only the cells which the expression of VEGF is high have the potential to form vasculogenic mimicry. But, it doesn't play a unique key role in vasculogenic mimicry.