Abstract: Objective To investigate the role of cytoskeletal-associated protein 2 （CKAP2） in human retinal vascular endothelial cells cultured in high glucose. Methods Human retinal microvascular endothelial cells （HRMECs） were cultured in vitro and divided into normal control group （CON group）, hypertonic culture group （30 mmol/L mannitol, HO group）, and high glucose culture group （30 mmol/L D-glucose, HG group）. Real-time PCR and Western blot were used for detecting the mRNA and protein expression level of CKAP2. After transfection of HRMECs with siCKAP2 to block CKAP2, or addition of Ranibizumab（106 μg/mL） to block VEGF, MTT, scratch and tubule formation experiments were performed to detect the proliferation rate, migration ability and tube formation ability of cells. Results ①Compared with the CON group and HO group, the mRNA and protein expressions of CKAP2 in the HG group were up-regulated; ② After siCKAP2 transfection, the cell proliferation ability, migration ability and angiogenesis ability were significantly reduced; ③Blocking CKAP2 and blocking VEGF have similar effects on reducing cell abilities in proliferation, migration, and tubular formation. Conclusions CKAP2 may be involved in the proliferation, migration and tube formation of retinal microvascular endothelial cells, and its effect may be similar to VEGF.