Inhibitory Effect Of Enoxacin On Osteosarcoma 143B Cells And Xenograft Tumors

Objective:To investigate the effects of enoxacin on the proliferation,invasion and apoptosis of osteosarcoma 143B cells and on the volume of xenograft tumor in mice.Methods:1.Osteosarcoma 143B cells were treated with different concentrations of enoxacin,and the untreated cells were used as control group.CCK-8 cell proliferation assay,Tumour-cell clonogenic assay,Transwell assays and Annexin V/propidium iodide(PI)apoptosis assay were used to detect the ability of proliferation,migration,invasion and apoptosis of osteosarcoma 143B cells.The protein expression levels of Bcl-xL,Bxl-2,MMP2 and MMP9 were analyzed by Western blot.2.100ul of 143B cell suspension(1×10~7 cells/ml)was injected into the tibial plateau of nude mice.The mouse xenograft tumor model was established and intraperitoneally injected with different concentrations of enoxacin(diluted with normal saline).The uninjected treatment group was used as the control group,and the tumor volume and body weight of mice were measured every week.Four weeks later,the heart blood of anesthetized mice was taken for liver and kidney function and blood cell test.To evaluate the hematological side effects of enoxacin in mice.After euthanasia,the tumor was dissected and its weight and volume were measured,and the differences among groups were observed.Results:1.CCK-8 cell proliferation assay,Tumour-cell clonogenic assay,Transwell assays and Annexin V/propidium iodide(PI)apoptosis assay showed that enoxacin could inhibit the proliferation,invasion and migration of 143B cells and induce their apoptosis(P<0.05).2.Western blot analysis showed that the expression of MMP2,MMP9,Bcl-xL and Bxl-2 protein in 143B cells decreased after enoxacin treatment,indicating that the down-regulation of MMP2 and MMP9 protein expression in 143B cells by enoxacin may be an effective way to inhibit osteosarcoma metastasis,while down-regulation of Bcl-2 and Bcl-xL protein expression may trigger the mitochondrial apoptosis pathway in osteosarcoma 143B cells.3.The volume and weight of tumor in mice treated with enoxacin were significantly lower than those in the control group(P<0.05),but there was no significant difference between the normal saline group and the control group(P?0.05).During the study period,there was no significant difference in body weight among the groups(P?0.05).These data suggest that enoxacin may have an inhibitory effect on the growth of osteosarcoma and has no obvious drug toxicity in nude mice.4.In the detection of liver and kidney function of heart blood of mice,no abnormality of liver and kidney function was found in each group after treatment with enoxacin,and the red blood cell,white blood cell and platelet count of mice were all in the normal range.The results showed that enoxacin had no obvious hematological side effects in the dose range of this experiment.Conclusion:Enoxacin inhibits proliferation,invasion and promotes apoptosis of osteosarcoma 143B cells.In animal experiments,enoxacin reduced the volume of xenograft tumor in mice.Enoxacin may be a promising new drug in the treatment of osteosarcoma,which is worthy of further study in clinical research.