【Abstract】 Objective To observe the role and effect of Src on anoikis in osteosarcoma. Methods Ultralow attachment 6 well plates were applied to establish the anoikis resistance model of osteosarcoma.Western blot assays were used to detect the expression of total Src and phosphorylated Src. After treated with Src inhibitor, the levels of Src and phosphorylated Src were tested, and the apoptosis rate was examined by using TUNEL assays and flow cytometry. Results The anoikis resistance model of osteosarcoma was successfully established. The phosphorylated Src′s gradual upregulation was accompanied with gradual enhanced anoikis resistance. The expression of phosphorylated Src in anoikis resistance osteosarcoma cells was higher than that of wide type (P＜005). After treated with Src inhibitor (PP2), phosphorylated Src, the active form of Src, was significantly repressed, while the apoptosis was clearly restrained (P＜005). Conclusion Src regulates anoikis in osteosarcoma can inhibite tumor cell apoptosis and contribute to its anoikis resistance.