基于多组学整合分析瞬时受体电位通道在肝癌发生发展中的分子分型及机制OA

Objective To investigate the biological mechanism and therapeutic potential of transient receptor potential(TRP)channels in the occurrence and development of hepatocellular carcinoma(HCC).Methods Differential genes related to TRP pathway were screened by integrating the TCGA,MsigDB,KEGG and Genecards databases,and molecular subtypes were identified by unsuper-vised clustering.A prognostic model was constructed using univariate Cox and LASSO regression,and validated through Bootstrap resampling(1 000 times)and external cohorts.The CIBERSORT algorithm was employed to evaluate the degree of immune cell infiltration,and the correlation between TRP prognostic risk score(TRPRS)and expressions of immune checkpoint molecules was analyzed.The clinical relevance of the prognostic model was assessed by drug sensitivity prediction combined with copy number variation analysis.The mRNA transcription levels of key genes(GHR,KCNJ11)were detected in HCC cell line MHCC-97H and normal hepatocyte line LO2 by qRT-PCR,and the protein expression levels of two key genes in HCC cells were measured using Coomassie brilliant blue staining and validated.Results A total of 93 TRP-related differentially expressed genes were identified and categorized into two molecular sub-types(cluster 1 and cluster 2)based on unsupervised clustering.The core genes GHR and KCNJ11 were selected by LASSO-Cox re-gression to construct a prognostic model,and the reliability of the model was confirmed by ROC curves in both internal and external co-horts(AUC>0.7,P=0.000 14 in TCGA_HCC training cohort).Multivariate Cox analysis indicated that TRPRS was an independent prognostic factor in TCGA_HCC(P<0.001),GSE76427(P=0.045),and LIRI-JP(P=0.03)cohorts.Additionally,analyses of immune infiltration and immune checkpoints revealed that TRPRS was associated with immune microenvironment characteristics(positively cor-related with CTLA4,negatively correlated with CD274,both P<0.05).Drug sensitivity analysis demonstrated that the patients with high TRPRS had lower TIDE score(P=0.006 7),and the sensitivity to chemotherapeutic drugs such as cisplatin increased in high TRPRS group(P<0.001).Results of qRT-PCR and Coomassie brilliant blue staining showed that both the mRNA(P<0.001)and total protein expression levels of GHR and KCNJ11 were significantly higher in HCC cell line MHCC-97H than in normal hepatocyte line LO2.Conclusion The TRP pathway factors GHR and KCNJ11 are involved in the progression of HCC by regulating the immune microenvironment and drug sensitivity,which provides a basis for mechanism research and therapeutic target screening of HCC.