Aim To observe the effect of miR-206 on proliferation of vascular endothelial cell and its mechanism, and provide a new therapeutic target for clinical arteriosclerosis. Methods The arterial endothelial tissue on lesion site and normal endothelial tissue were collected from the patients who were diagnosed as diabetic lower limb atherosclerosis by first time in clinical pathology. The expression of miR-206 was detected by qPCR. Normal HUVEC cell lines, HUVEC transfected with miR-206-mimics and HUVEC transfected with miR-206-inhibitor were studied through fundamental experiment, and the transfection efficiency were observed. In the three groups, CCK8 was used to detect the proliferation status, transwell was used to evaluate the migration ability, flow cytometry was used to detect cell cycle and Western blot was used to observe expression of Cx43. Results The mRNA expression of miR-206 on lesion site of lower limb artery (shin peroneal artery) in the patients with diabetic arteriosclerosis was significantly increased compared with normal endothelial tissue(P<0.05); The expression of miR-206 significantly increased (P<0.05) after transfected with miR-206-mimics, while it was significantly reduced (P<0.05) after transfected with miR-206-inhibitor. Compared with normal HUVEC and HUVEC transfected with miR-206-inhibitor, the cell proliferation rate was significantly lower in HUVEC transfected with miR-206-mimics for 24 h (P<0.05). After HUVEC was transfected with miR-206-mimics, the cell migration capacity was significantly lower than normal HUVEC group (P<0.05), and the cell cycle was blocked during G2 phase; while the cell migration capacity was higher in HUVEC transfected with miR-206-inhibitor group than that of normal HUVEC group (P<0.05). Cx43 expression was significantly reduced in miR-206-mimics group but significantly increased in miR-206-inhibitor group. Conclusion miR-206 can inhibit the proliferation of vascular endothelial cells, block endothelial cells in G2 phase and suppress their migration, and it may affect proliferation of endothelial cells by regulating Cx43 expression.