Objective The mechanism of inhibition of the proliferation and migration of osteosarcoma cells by miR-646 was investigated.Methods The effects of miR-646 on the proliferation of U2OS cells were detected by MTT assay.The effects of miR-646 on U2OS cell migration were analyzed by Tmnswell assay.MIRDB was used to predict the binding sites of miR-646 and epidermal growth factor receptor (EGFR),which were confirmed in a luciferase reporter assay.Western blotting was conducted to detect the effect of miR-646 on the EGFR pathway.Results The MTT test showed that transfection of miR-646 significantly inhibited U2OS cell proliferation;the 24 h inhibition rate was 21.76％± 1.05％.In contrast,the 24 h promotion was 18.89％± 0.81％.Transwell experiments showed that miR-646 inhibited the migration of osteosarcoma U2OS cells.MIRDB predicted that miR-646 bound in the 3'-untranslated region of EGFR,and luciferase reporter gene experiments showed that miR-646 directly acted on EGFR.Western blotting experiments demonstrated that miR-646 inhibited the proliferation and migration of U2OS cells to some extent by inhibiting the EGFR pathway.Conclusion miR-646 can inhibit the proliferation and migration of osteosarcoma cells by inhibiting the EGFR pathway.