圣和散激活FoxO3a增强人恶性胶质母细胞瘤放疗敏感性的机制OA

Objective:To investigate the mechanism of Shenghe Powder(SHP)in enhancing the radiotherapy sensitivity of human malignant glioblastoma U251 cells through the FoxO3a signaling pathway.Methods:U251 cells were taken as the research subject,and the normal control(Control)group,SHP group,negative control(SHP+si-NC)group,and knockdown FoxO3a(SHP+si-FoxO3a)group were set up,and the cells were irradiated with a single dose of 5 Gy 6 MV X-ray.Western blot,immu-nofluorescence staining,CCK-8,Edu,flow cytometry,and cancer stem cell spheroidization were used for detection.Through es-tablishing a tumor-bearing nude mouse model,tumor changes were detected,and tumor tissue-related proteins were detected by immunohistochemistry.Results:Compared to the Control group,the expressions of BNIP3 and FoxO3a proteins of cells in the SHP group were increased,and the expression of p-FoxO3a protein was decreased(P<0.01).The red fluorescence of FoxO3a in the SHP group cells was strongly expressed in the nucleus(P<0.001).The green fluorescence of BNIP3 was strongly expressed on the mitochondrial membrane in the SHP group cells(P<0.001).The cell viability,positive cell rate,number of cell migration and invasion,and spheroidal number of cells in the SHP+si-FoxO3a group were significantly higher than those in the SHP+si-NC group,and the apoptosis rate was significantly lower than that in the SHP+si-NC group(P<0.01).The expressions of Cl-caspase3,Beclin1,LC3Ⅱ/LC3Ⅰ,BNIP3,and FoxO3a proteins in the SHP+si-FoxO3a group cells were significantly lower than those in the SHP+si-NC group,and the expression of p-FoxO3a protein was significantly higher than that in the SHP+si-NC group(P<0.01).Tumors grew slowly in the SHP group compared to the Control group;Tumor growth was slower in the SHP+si-FoxO3a group compared to the SHP+si-NC group.Compared to the Control group,the volume and weight of tumors in the SHP group were significantly reduced;Compared to the SHP+si-NC group,the tumor volume and weight of the SHP+si-FoxO3a group were significantly increased(P<0.01).Compared to the Control group,the tumor tissue expression of Cl-caspase3,Beclin1,and BNIP3 proteins in the SHP group was significantly increased(P<0.01),and the expression of p-FoxO3a protein was significantly decreased(P<0.01).Compared to the SHP+si-NC group,the tumor tissue expression of Cl-caspase3,Beclin1,and BNIP3 proteins in the SHP+si-FoxO3a group was significantly reduced(P<0.01),and the expres-sion of p-FoxO3a protein was significantly increased(P<0.001).Conclusion:Cellular experiments shows that SHP enhances the FoxO3a-BNIP3 signaling pathway by inducing FoxO3a dephosphorylation to the nucleus,and BNIP3 translocation to the mito-chondrial membrane.Knockdown of FoxO3a reverses the enhanced effect of SHP on the radiosensitivity of glioma U251 cells.In vivo experiments also confirm that SHP enhances the radiotherapy sensitivity of human malignant glioblastoma U251 cells through the FoxO3a-BNIP3 signaling pathway.