The Role Of RIPK3 In Prognosis Of Patients With Low-grade Glioma

Objective: Preliminary explore necrotizing apoptosis related gene in patients with low grade glioma(LGG)and prognostic significance of expression,and to verify the expression and localization of key genes.Methods: The sequencing data and clinical data of 515 LGG patients,including gender,M stage,Grade grade and survival time,were downloaded from the TCGA database,and the necrotic apoptosis related genes were retrieved from the GSEA database.Univariate and multivariate COX regression analyses were used to screen necrotic apoptosis related genes associated with prognosis,and a prognostic risk scoring model was constructed by LASSO regression to divide the LGG patients into high-risk groups and low-risk groups,and to evaluate the effect of LGG prognostic related genes in distinguishing high and low risk groups.Kaplan-Meier(K-M)survival analysis was used to evaluate the high-risk group and the low-risk group.Time-dependent offset operating characteristic curve(ROC)was used to evaluate the model prediction effect.Through the enrichment of GO and KEGG pathways in high and low risk groups,the possible causes of risk differences were explored.Finally,the immune function status of high and low risk groups was investigated by ss GSEA analysis.Collected in our department surgery and postoperative pathological diagnosis of low-grade gliomas,for the second time diagnosed with recurrent or worsening of LGG patients with complete data of medical records,incorporated in the recurrent group.In addition,patients with low-grade glioma confirmed by surgery and postoperative pathology,but not yet recurrent,were included in the control group.The wax blocks of the patients’ pathological tissue were collected,and immunohistochemical(IHC)staining was performed to verify the expression and localization of the gene,which related to necrotic apoptosis of LGG.Results: 7 necrotic apoptosis genes related to prognosis in LGG patients were identified by COX regression analysis,FADD,FAS,MLKL,RIPK1,RIPK3,TLR3 and TNF,among which RIPK3 had the strongest correlation with other genes and was used in subsequent IHC staining.In the LASSO regression model,7 genes were distributed in two directions at different risk components,indicating that the prognostic risk score model of these 7 genes could distinguish the high-risk group from the low risk group.Kaplan-Meier(K-M)survival analysis and ROC curve showed that the overall survival rate of the high-risk group was lower than that of the low-risk group.KEGG and GO enrichment analysis showed significant differences in immunity between the high-risk and low-risk groups,while ss GSEA analysis scores showed significant differences in scores of most immune cells between the high-risk and low-risk groups,with the high-risk group scoring significantly higher than the low-risk group.Immunohistochemical staining of RIPK3 expression in focal tissues showed that there were significant differences in the expression of RIPK3 between the relapsed group and the control group.Conclusion:Seven genes,FADD,FAS,MLKL,RIPK1,RIPK3,TLR3 and TNF,were positively correlated with necrotic apoptosis in LGG patients,among which RIPK3 was involved in necrotic apoptosis in the course of low-grade glioma,and was highly expressed in lesion tissues of low-grade glioma recurrence patients,suggesting bad prognosis.