Expression,Role And Prognostic Value Of POFUT1 In Glioma

Objective: To investigate the expression and biological function of O-fucosyltransferase 1(POFUT1)in glioma,and to preliminarily explore the molecular mechanism by which POFUT1 exerts its regulatory effect on glioma.Methods: 1.CGGA database and TCGA database were used to analyze the expression of POFUT1 in glioma tissues.Chi-square test was used to analyze the correlation between the expression of POFUT1 and the clinical information of patients.Kaplan-Meier survival analysis and COX regression analysis were used to analyze the survival prognosis between the expression of POFUT1 and the clinical information of patients;2.The expression level of POFUT1 in different grades of gliomas was tested by immunohistochemistry,and the relationship between the expression level of POFUT1 and clinicopathological parameters was studied by chi square test,Kaplan Meier survival analysis and COX regression analysis;3.U251 cell line stably overexpressing POFUT1 was constructed by lentivirus transfection technology,and knock down POFUT1 gene in U251 and U87 cell lines by si RNA.The protein expression level and m RNA expression level of POFUT1 were detected by Western blot and Real time quantitative polymerase chain reaction(RT q PCR);The effects of POFUT1 on the proliferation,invasion and migration of glioma cells were detected by MTT assay and Transwell assay;4.Xenograft tumor model was used to validate the effect of POFUT1 on the proliferation of glioma;5.We performed the transcriptome of POFUT1 overexpression cell lines and knockdown cell lines,screened out the significantly differentially expressed genes,and analyzed the enrichment of Kyoto Encyclopedia of genes and genomes(KEGG)and gene set enrichment analysis(GSEA).Results: 1.The expression level of POFUT1 in glioma tissues was significantly higher than that in normal tissues adjacent to the tumor(p < 0.05);the prognosis of patients with high expression of POFUT1 glioma was significantly worse than that of patients with low expression(p < 0.001);2.POFUT1 expression status was correlated with patients’ age,gender,onset status,clinical stage,and IDH mutation type;in different age groups,different genders,LGG group,GBM group,IDH mutation group,IDH wild group,primary group,recurrence group,MGMT methylation group,and MGMT non-methylation group,the survival of patients with high expression POFUT1 was significantly higher than those of low-expressing patients(p < 0.05);a prognostic model that takes into account POFUT1 factors can better assess the prognosis of patients;3.Overexpression of POFUT1 promoted glioma cell migration and invasion,and knockdown of POFUT1 inhibited glioma cell migration and invasion(p < 0.001);4.Overexpression of POFUT1 promoted subcutaneous tumorigenesis in nude mice(p < 0.001);5.Transcriptome sequencing after POFUT1 overexpression and knockdown revealed significantly different genes mainly enriched to JAK/STAT,ECM receptor,TGF-β,P53,PI3K/AKT,cell adhesion,cell cycle and other pathways.Conclusion: POFUT1 can promote the proliferation and invasion of glioma,and can preliminarily evaluate the prognosis of glioma.