Investigation On The Effect Of TBC1D15 On Proliferation And Apoptosis In Glioblastoma

Background: Glioma is the most common primary intracranial malignant tumor.Glioblastoma accounts for 54% of all gliomas and 60-70% of high-grade gliomas.The survival time for low-grade glioma is 5-10 years,for anaplastic astrocytoma 2-3 years,and for glioblastoma only 12-15 months.The 5-year survival rate of patients with glioblastoma is as low as 3%.Although glioma is treated in a variety of ways,including surgical resection,radiotherapy,chemotherapy,as well as emerging immunotherapy and electric field therapy,the treatment effect of glioma is still poor,and the studies over the past few decades have only extended the median survival time with a few months.Glioma has always been the focus and difficulty of central nervous system tumor research.Objective: By collecting tissue samples from glioma patients and using Western Blot to detect the expression of TBC1D15 protein in glioma tissue samples,we can understand its differential expression in normal brain tissue and glioma and further investigate the effect of TBC1D15 on the proliferation and apoptosis and its potential mechanism in U87 glioma cells.Methods:1.24 glioma specimens and 5 normal brain tissues were collected from the department of neurosurgery,affiliated hospital of Qingdao university from December 2018 to April 2019.the expression of TBC1D15 was detected by western blot.2.Detection of siRNA interference: the expression of TBC1D15 in U87 cells was knocked down by siRNA transfection,and the interference efficiency of siRNA was detected by Western Blot.3.Proliferation test: U87 cells were transfected with TBC1D15-siRNA and control-siRNA,the proliferation of U87 cells in the TBC1D15-siRNA group and Control-siRNA group for 24 h,48h,72 h was detected by CCK8,respectively.4.Apoptosis test: U87 cells were transfected with TBC1D15-siRNA and control-siRNA,flow cytometry detected the apoptosis of U87 cells in the TBC1D15-siRNA group and Control-siRNA group.5.Wnt/β-catenin signal pathway test: U87 cells were transfected with TBC1D15-siRNA and control-siRNA,and then cellular protein was extracted.western blot tested the expression of β-catenin in U87 cells in the TBC1D15-siRNA group and Control-siRNA group.Results:1.The expression level of TBC1D15 in glioma tissues and normal brain tissues was tested by Western Blot,indicating that the expression of TBC1D15 in glioma tissues was significantly higher than that in normal brain tissues(P < 0.05),and the expression of TBC1D15 in glioma tissues was correlated with WHO grading.2.The proliferation of U87 cells was analyzed by CCK8 assay,indicating that the proliferation in the TBC1D15-si RNA group was significantly lower than that in the Control-siRNA group(P < 0.05).3.The apoptosis of U87 cells was assayed by flow cytometry,showing that the apoptosis rate in TBC1D15-si RNA group was significantly higher than that in Control-siRNA group(P < 0.05).4.Western blot assay tested the expression of β-catenin in U87 cells,indicating the expression of β-catenin in the TBC1D15-si RNA group was significantly lower than that in the Control-siRNA group(P < 0.05).Conclusion: The expression level of TBC1D15 in glioma tissues was significantly higher than that in normal brain tissues,and its expression level was positively correlated with the WHO grade,that is,the higher the WHO grade of glioma,the higher the expression of TBC1D15.TBC1D15 Knockdown in U87 glioma cells via transfecting siRNA can inhibit the proliferation of glioma cells and promote the apoptosis of glioma cells,and the mechanism may be to inhibit the activation of Wnt/β-catenin signaling pathway.