Expression And Effects Of T Cell Immunoglobulin And Mucin Domain Containing Molecule 3 In Osteosarcoma

Part ?Expression of TIM-3 in the osteosarcoma and analyze of the clinical significancesObjective:To test the expression levels of TIM-3 in osteosarcoma,and analyze the correlation between expression level and the clinical prognosis of patients.Methods:Application of immunohistochemical technique detection of 38 cases of osteosarcoma in tumor tissue and tissue adjacent to carcinoma to test TIM-3 expression level,and analyze the correlation between expression level and the clinical prognosis of patients by using statistical method.Results:Immunohistochemical results show that the TIM-3 in osteosarcoma tissues present high expression of different level,low expression in the tissue adjacent to carcinoma.There is no statistical differences between TIM-3 expression level of patients with osteosarcoma and gender,age,tumor pathologic type,pathological changes,Enneking stages.Kaplan and Meier survival analysis show that TIM-3 clinical outcomes for patients with high expression levels of group is poorer than the low expression level group.Conclusion:Expression level of TIM-3 in osteosarcoma tumor tissue is higher,but the expression in the tissue adjacent to carcinoma is lower,clinical outcomes for osteosarcoma patients with high expression level of TIM-3 is poor than low negative expression level,TIM-3 may play an important role in the osteosarcoma development.Part ?Inhibiting effects of biology behavior on osteosarcoma cell by targeting TIM-3Objective:To study osteosarcoma cell proliferation,apoptosis,migration,invasion and other biological behaviour by targeting TIM-3,and to provide theoretical basis for TIM-3 immunotherapy.Methods:Application of MTT and clone forming experiment testing to determin proliferation inhibition of osteosarcoma MG63 cells and U-20S by targeting TIM-3;Application scratch experiment and Transwell detection to determin inhibition on osteosarcoma cell migration,invasion by targeting TIM-3;Application of flow cytometry instrument Annexin V-FITC staining method to detect influence on osteosarcoma cell apoptosis by targeting TIM-3;Using Western blot technique to detect invasion related protein expression levels on osteosarcoma cells by targeting TIM-3.Results:MTT results suggest targeting inhibition of TIM-3 can significantly inhibit proliferation and clone formation of osteosarcoma MG63,U-20S;Scratch experiment,Transwell experiment suggests that targeting inhibition of TIM-3 could inhibit the MG63,U-20S migration and invasion;Flow cytometry instrument Annexin V-FITC staining results suggest targeting inhibition of TIM-3 can promote MG63,U-20S apoptosis;Western blot results suggest targeting TIM-3 can significantly reduce the invasion related protein expression.Conclusion:Targeting inhibition of TIM-3 can inhibit osteosarcoma cell proliferation,migration and invasion,and promote apoptosis.Part ?Antitumor effect by targeting TIM-3 of the osteosarcoma tumor-burdened nude miceObjective:To test antitumor effect(weight change and tumor volume)on osteosarcoma U-20S tumor-burdened nude mice.Methods:24 osteosarcoma U-20S tumor-burdened nude mices were divided into TIM-3 siRNA group and control group,the U-20S cells were set into tumor-burdened nude mice model in right back subcutaneous,to test nude mice weight change and tumor volume and then drawed growth curve,when the first nude mouse normal death we killed all the nude mice,and tested weight and volume of the tumor.Application of immunohistochemical techniques to test the expression levels of PCNA and HMGB1 in the tumor tissue of both TIM-3 siRNA group and control group.Results:TIM-3 siRNA group significantly reduce the tumor weight and volume and inhibitory rate increased significantly compared to the control group.PCNA and HMGB1 expression levels in experimental TIM-3 siRNA U-20S tumor-burdened nude mice group were significantly lower than the control group.Conclusion:siRNA targeted TIM-3 can inhibit osteosarcoma cells growth and invasion ability in osteosarcoma U-20S tumor-burdened nude mice.TIM-3 may play an important role in the osteosarcoma development.