Effect Of Indomethacin On Proliferation,Apoptosis,Migration And Invasion Of Osteosarcoma And Its Mechanism

Objective: Osteosarcoma(OS)is a common malignant bone tumor that occurs in adolescents and children.The current gold standard of treatment is preoperative chemotherapy plus surgical treatment.But tumor resistance limits the use of chemotherapy drugs,and the prognosis remains poor.Indomethacin is a non-steroidal anti-inflammatory drug widely used to treat inflammation.In recent years,it has shown antitumor effects in a number of tumors.However,the efficacy of indomethacin in the treatment of osteosarcoma is still unclear,and the mechanism of action needs further study.In this study,we examined the effect of indomethacin on proliferation,apoptosis,migration and invasion of osteosarcoma and its molecular mechanism.Methods: Osteosarcoma U2 OS cells were cultured in 5A medium of Mc Coy.Cell proliferation were measured by CCK8 assay and colony-formation assay.Cell death was detected using Hoechst 33258 staining kit and annexin V fitc-pi apoptosis kit The Transwell test is also used to detect cell invasion and migration.Last,the expression levels of Bax,Bcl-2,P-AKT,Total-AKT,β-catenin and MMP-2 were analyzed by western-blot analysis.Results: CCK-8 assay and colony-formation assay showed that indomethacin inhibited the proliferation of osteosarcoma U2 OS cells compared with the control group,and the proliferation ability of the cells gradually decreased with the increase of concentration.Flow cytometry detection,in addition,Hoechst 33258 staining and western blot showed that indomethacin induced osteosarcoma U2 OS cells apoptosis by adjusting the Bax and Bcl-2 protein.the expression of Bcl-2 was decreased and expression of Bax have no obvious change.In addition,transwell assay shows that indomethacin can inhibit invasion and migration of osteosarcoma U2 OS cells by decreased the expression of MMP-2.Last,western blot showed that compared with the control group the expression of P-AKT and β-catenin were decline;There was no significant change in the levels of Total-AKT and,suggesting that indomethacin may regulate the phenotype of osteosarcoma U2 OS cells through the AKT /β-catenin signaling pathway.Conclusion: Indomethacin can inhibit the proliferation,invasion and migration of osteosarcoma U2 OS cells and lead to cell apoptosis.With the increase of indomethacin concentration,the ability to inhibit the proliferation,invasion,migration and induction of apoptosis of osteosarcoma U2 OS cells increases gradually.The potential mechanism of indomethacin against osteosarcoma U2 OS cells may be through the regulation of AKT/-catenin signaling pathway...