Oral Squamous Cell Carcinoma M6A Was Constructed Based On The TCGA Database Prognostic Risk Model For Methylation Factors

Objective: The purpose of this study was to establish and apply the prognostic model of m6 A regulator in oral squamous cell carcinoma and preliminarily investigate the mechanism of m6 A regulator.Methods: The data set of oral squamous cell carcinoma patients included RNA sequencing data,methylation data,and clinical information obtained from the TCGA database,and the m6 A gene and its validated target genes were obtained from the RM2 TARGET database.A new rank based pairwise comparison algorithm was innovatively adopted to select effective m6 A gene pairs,which were defined as m6 A regulators.The patients obtained from the TCGA database were randomly divided into the training group(2/3)and the test group(1/3).In the training set,the collinearity of the m6 A regulator was removed by Lasso-Cox regression.Multivariate Cox regression analysis was used to construct prognostic model.Receiver operating characteristic(ROC)curve,C index,calibration curve,cumulative risk curve,and independent prognostic analysis were used in model validation to verify the predictive performance of prognostic model survival.Apply the same analysis to the test set.To explore the potential mechanisms of the occurrence and prognosis of oral squamous cell carcinoma by using a comprehensive GSEA algorithm.In addition,we described the mutant landscape of genes in high-and low-risk groups and prognostic models of oral squamous cell carcinoma,identifying key genes.The key gene mechanisms were investigated from RNA mutation,protein expression in HPA database,TISCH database single cell analysis,and multi-software immunocorrelation analysis,and verified by immunohistochemistry.Prognostic model was used to predict the sensitivity of oral squamous cell carcinoma to chemotherapy and immunotherapy.Finally,the traditional clinical variables were combined to construct a histogram to visualize the prognostic model.Results:(1)6-m6 A regulator: ELAVL1 | LMNB1,YTHDF2 | YAP1,ELAVL1 | VEGFA,YTHDF1 | CDK4,WTAP | CDKN2 A and YTHDF2 | GADD45 A.(2)Single-Multivariate COX analysis showed that risk score was an independent predictor of poor prognosis for OSCC(HR:1.9,P<0.001).The prognosis model was constructed based on the 6-m6 A regulator and the visualization of the column graph.The survival curve showed that the low-risk group had a better prognosis.ROC curve shows that(AUC1:0.725;AUC3:0.753;AUC5:0.747),6-m6 A regulators have good predictive performance.(3)Gene function enrichment analysis showed that the high-risk group was mainly enriched in immune and cell cycle related signaling pathways,while the low-risk group was mainly enriched in cell differentiation related signaling pathways.(4)Mutation analysis suggested that high and low risk groups had similar mutation patterns in genes,protein domains,and cell signaling pathways.(5)CDKN2A was the gene with the highest mutation frequency in the model(up to 20%)and was highly expressed in the low-risk group(P=0.006).The gene with the highest deletion frequency was CDKN2A(more than 30%),but GSEA suggested that the CDKN2 A mutant group had better immune function.HPA database and immunohistochemistry suggested that CDKN2 A was positive in oral squamous cell carcinoma(P<0.001),and patients with high CDKN2A/p16INK4 a expression had better prognosis(P=0.038).(6)Single-cell analysis: GSE139324 suggested that CDKN2 A was highly expressed in T cells,GSE172577 suggested that CDKN2 A was highly expressed in duct cells and glandular cells,and GSE103322 suggested that CDKN2 A was significantly enriched in malignant cells.(7)Tumor immuno-phenotype tracking(TIP)ss GSEA score suggested that the high-risk group had more immune reserve.Immunophenotyping(IPS)and tumor Immune dysfunction and rejection(TIDE)scores suggested that high-risk groups were more suitable for immunotherapy.(8)Cmap database analysis suggested that imatinib was the drug most likely to reverse the status of the high and low risk group(score =-1).Conclusion: This study is helpful to further understand the role of 6-m6 A regulators in the prognosis and development of oral squamous cell carcinoma,and provides a theoretical basis for future immunotherapy and chemotherapy drug research.