| Background:Ewing’s sarcoma is the second most common malignancy of bone and soft tissue in children and adolescents.The incidence rate is 1 per 1.5 million people,which is higher in the population of European origin and slightly biased in males.Tumors can occur in any part of the body,but the pelvis and proximal long bones are most often involved.The clinical features of Ewing’s sarcoma are largely non-specific.Therefore,early patients are very easy to cause misdiagnosis,missed diagnosis,delay the disease,leading to the development of the disease,which is a catastrophic consequence for patients.The tumor necrosis factor-α-induced protein 8-like protein 1 is a newly discovered protein involved in the death domain of immune regulation and tumorigenesis.After decades of research,it has been found that this protein plays an important role in immune homeostasis,inflammation and cancer development regulation.And it plays the role of tumor suppressor in several kinds of cancer.Previous studies have shown that activation of the Wnt/β-catenin signaling pathway in tumor cell subpopulations promotes phenotypic heterogeneity and disease progression in Ewing’s sarcoma.Nevertheless,the exact role of TIPE1 in Ewing’s sarcoma remains ambiguous.Purpose:The purpose of this study is to explore the biological behavior of Ewing’s sarcoma and to further explore the expression,role and related mechanism of TIPE1 in Ewing’s sarcoma.Methods:In the present study,TIPE1 expression in Ewing’s sarcoma cells was determined by q PCR and western blotting.Further,Ewing’s sarcoma cell line RD-ES was transfected with lentivirus-based TIPE1 expression system to upregulate the expression of TIPE1.The effects of TIPE1 on the proliferation of Ewing’s sarcoma cells were detected by Cell Counting Kit-8(CCK-8).The effects of TIPE1 on the migration and invasion of Ewing’s sarcoma cells were detected by Transwell assay.The apoptosis rate of Ewing’s sarcoma cells was detected by flow cytometry,and the apoptotic protein level was detected by Western blotting.The expression levels of key proteins in the Wnt/β-catenin signaling pathway were detected by Western blotting to clarify the mechanism of action of TIPE1 on Ewing’s sarcoma.Results:Studies have shown that compared with human normal cells,the m RNA level of TIPE1 in Ewing’s sarcoma cells is significantly down-regulated,and the protein expression level of TIPE1 is also down-regulated.The difference of its expression was statistically significant.In the migration and invasion experiment,the overexpression of TIPE1 significantly inhibited the migration ability of RD-ES cells,and also led to a significant decrease in the invasive ability of RD-ES cells.In the detection of flow cytometry,it was found that the apoptosis rate of TIPE1 overexpression group was significantly higher than that of control group.Then the expression level of apoptosis-related proteins was detected by Western blotting,and it was found that the change of the expression level of apoptosis-related proteins and regulating the expression of apoptosis-related proteins was one of the mechanisms of inducing apoptosis.By measuring the expression level of key proteins in Wnt/β-catenin signal pathway,it is confirmed that the overexpression of TIPE1 can inhibit Wnt/β-catenin signal pathway,and then inhibit the growth,movement and survival of Ewing sarcoma.Conclusion:In summary,our results demonstrate the anti-tumor function of TIPE1 in Ewing’s sarcoma.These findings may help doctors better understand the pathogenesis and development of Ewing’s sarcoma and reveal a novel therapy target for Ewing’s sarcoma. |
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