Vascular endothelial growth factor (VEGF), one of the most important angiogenic factor, can impact the tumor cell proliferation and invasion, but the mechanism remains unclear. This study is to investigate the key proteins which may play an important role in the VEGF-induced progress of ovarian cancer cells. The total protein from HO-8910 cells was separated by two-dimensional electrophoresis (2-DE), and differentially expressed proteins were identified by matrix-assisted laser desorption and ionization time-of-flight tandem mass spectrometry (MALDI-TOF MS) and PDQuest image analysis software. Furthermore, real-time PCR, Western blot, and immunocytochemistry were also used to confirm different expression levels of differential proteins. Morphological changes and invasion capability were evaluated by electron microscope and Matrigel invasion assay, respectively. The highly reproducible and well-resolved 2-DE patterns of both HO-8910/VEGF and HO-8910 cells were acquired. A total of 17 expressed differential proteins were identified, 8 proteins were upregulated (ACTB, TIM, PDIA3, PDIA1, DCTN2, KIC17, SIAS, and KIC10) and 9 downregulated (KIC18, GRP78, CAPG, PPIA, ROA2, LMNA, EZRI, ADRM1, and ENOA). Ultrastructure of VEGF-treated group showed more malignant characteristic compared with control group, an obvious increase in the number of cells penetrating the Matrigel membrane in VEGF-treated group (P < 0.05). These results suggested that VEGF could impact ovarian cancer's malignant progression by regulating expression of associated proteins.