| Hepatocellular carcinoma(HCC)is among the most common types of cancers that threat the public health worldwide.N6-methyladenosine RNA methylation,which is the most common form of mRNA modification and correlated with cancer initiation and progression,is dynamically regulated by m6A RNA methylation regulators,including writers,erasers,and readers.However,the prognostic role of m6A RNA methylation regulators in hepatocellular carcinoma is poorly understood.This study aims to explore the roles of m6A RNA methylation regulators in hepatocellular carcinoma,and identify a risk signature and prognostic values of m6A RNA methylation regulators in HCC.1.The landscape of m6A RNA methylation regulators in HCC samples from TCGA and their function in HCC.The RNA-seq transcriptome data and corresponding clinicopathological information were obtained for HCC samples and normal samples from TCGA.And we found that 19 m6A RNA methylation regulators were abnormally expressed in liver cancer tissues.Consensus clustering analysis was performed on 19 m6A RNA methylation regulators to identify two clusters of HCC with distinct clinical outcomes and clinicopathological features.The principal component analysis examined the validity of the clusters.Chi-square analysis showed that the clusters were related to clinical stage,tumor pathological grade and survival status.Survival analysis found that the patients in cluster 2 had worse clinical outcomes.Then differential gene function enrichment and GSEA analysis of the two clusters revealed that m6A RNA methylation methylation regulators are significantly related to the biological process and signaling pathways of malignant progression of HCC.2.The identification and prognostic value of th Risk Signature modelWe screened 12 m6A RNA methylation regulators related to the survival prognosis of liver cancer patients by univariate COX regression analysis and had them analyzed in LASSO regression,which could be constructed risk signature model of HCC patients based on 6 m6A RNA methylation regulators,including YTHDF2,YTHDF1,IGF2BP3,KIAA1429,METTL3 and ZC3H13.The analysis of ROC curve indicated the well-prediction performances of riskscore signature.Chi-square analysis suggested that the riskscore signature were closely related to the clinicopathological features of HCC patients such as T stage,clinical stage,tumor pathological grade,and survival status.Multivariate COX regression analysis showed that the riskscore signature of patients could be be served as an independent prognostic factor in HCC.3.NCBI and GEPIA analysis of YTHDF1 expression landscape in normal human tissues and other tumors.Compared with other normal tissues,YTHDF1 expression is lower in human normal liver tissue,but high-expression in HCC by the NCBI and GEPIA analysis.The expression level of YTHDF1 is significantly related to the overall survival time of patients.4.Expression of YTHDF1 in patients with HBV-associated hepatocellular carcinoma and its prognostic value.We employed real-time quantitative PCR and immunohistochemical staining to analyze the expression of YTHDF1 in HBV-associated HCC tissues and their adjacent parts separately.The expression level of YTHDF1 were was much higher in tumor tissues than in normal counterparts.And meanwhile,the expression level of YTHDF1 was correlated with tumor size,the existence of microvascular invasion(MVI),the existence of portal vein tumor thrombosis(PVTT),TNM staging,tumor histological differentiation,BCLC staging.Patients with higher expression of YTHDF1 in HBV-associated HCC tissues had a shorter relapse-free survival(RFS)than that in lower YTHDF1 expression group.And multivariate COX regression analysis suggested that the high expression of YTHDF1 in HBV-associated HCC tissues was an independent prognostic factor of RFS in HCC patients.In conclusion,m6A RNA methylation regulators are crucial participants in the malignant progression of hepatocellular carcinoma and are potentially useful for prognostic stratification and treatment strategy development. |
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