Vascular endothelial growth factor receptor 2 (VEGFR2) is highly expressed in non-small cell lung cancer (NSCLC), and is the main target of antiangiogenesis therapy against this disease. However, there is limited evidence regarding its regulatory mechanism. Thus, elucidating the underlying mechanism of regulation of VEGFR2 is of great value to antiangiogenesis therapy. The colocalization of VEGFR2 and small ubiquitin-like modifier 1 (SUMO1) was detected through confocal microscopy. We examined the level of VEGFR2 SUMOylation in cells and rat tissues, and its effects on the angiogenesis signaling pathway (immunoprecipitation and western blotting), as well as the proliferation (Cell Counting Kit-8 assay) and migratory ability (cell scratch and Transwell assays) of NSCLC cells. Apoptosis was evaluated through Hoechst staining. VEGFR2 and SUMO1 are colocalized in the cytoplasm. VEGFR2 can be SUMOylated through combination with SUMO1 in cells and rat tissues, and the level of VEGFR2 SUMOylation in NSCLC is higher than that observed in healthy cells and tissues. Cell proliferation, migration, and the protein levels of phosphorylated-VEGFR2/phosphorylated-Akt/phosphorylated-extracellular signal-regulated kinase 1/2 (p-VEGFR2/p-Akt/p-ERK1/2) were significantly increased in NSCLC cells transfected with VEGFR2 K1270R versus those reported in cells transfected with VEGFR2 (wild-type). The levels of p-VEGFR2/p-Akt/p-ERK1/2 protein were significantly decreased in cells transfected with sentrin-specific protease 1-targeting small interfering RNA (siSENP1) versus those recorded in nontransfected controls. VEGFR2 SUMOylation may play an important role in antiangiogenesis therapy of NSCLC. The level of VEGFR2 SUMOylation may be a prognostic marker in patients with NSCLC.