Screening And Analysis Of Potential Biomarkers Associated With Congenital Biliary Dilatation Malignancy

Objective:To explore potential biomarkers associated with congenital biliary dilatation（CBD）malignancy based on m RNA high-throughput sequencing technology and to explore the molecular mechanisms involved.Methods:recorded In this study,9 cases each of children with CBD hospitalized from June to December 2021 and healthy check-ups during the same period were selected.m RNA high-throughput sequencing was performed on 18 serological samples from the experimental and control groups,first by extracting total RNA from their serological samples,and by mass-checking the sample RNA,building libraries for sequencing as well as evaluation to obtain reliable gene expression data.Then we visualized the differential genes by building the R language environment for Gene Ontology（GO）enrichment analysis,Kyoto Encyclopedia of Genes and Genomes（KEGG）pathway enrichment analysis.Finally,the values of P_adjand log₂Fold Change were combined,and the differential genes of five were selected as the key genes of this study after the relevant differential genes were searched and reviewed in the Gene Cards database with reference to the interactions between the relevant differential genes and other genes in the network.Finally,RT-q PCR was used to verify the m RNA expression levels of the key genes in the CBD group and the healthy control group,and the subject receiver operating characteristic（ROC）curves were plotted to evaluate the diagnostic performance of the hub genes.Results:A total of 1799 differential genes existed in the CBD and Normal groups,among which 96 oncogenes were associated with CBD malignancy,including 56 up-regulated genes and 40 down-regulated genes.GO enrichment analysis showed that differential gene expression products played important roles in cellular composition of the cytoplasmic side of the plasma membrane,heterotrimeric G protein complex,GTPase complex,plasma membrane signaling receptor complex,etc.In terms of molecular function,differential genes were significantly enriched in protein heterodimerization activity,cytokine receptor binding,transcriptional coregulator binding,and transcriptional co-regulator binding,etc.KEGG pathway enrichment analysis revealed that differential genes were significantly enriched in transcriptional dysregulation in cancer,PI3K-Akt signaling pathway and pathways associated with human cytomegalovirus infection.Combining the values of P_adjand log₂Fold Change,and referring to the interactions between the relevant differential genes and other genes in the network,five differential genes（TRAF4,LPAR6,GLI1,CAV2,ATF1）were selected as hub genes for this study after searching and reviewing the relevant genes in the Gene Cards database.Finally,the results of RT-q PCR,ROC curve and literature search showed that LPAR6,CAV2 all had good diagnostic value.Conclusion:1.The relative m RNA expression levels of TRAF4,LPAR6,GLI1,CAV2,and ATF1genes were statistically significant in the CBD group and the healthy control group.Among them,the RT-q RNA of TRAF4,LPAR6,GLI1,CAV2 genes were consistent with the RNA-seq prediction.2.LPAR6,CAV2 are expected to be potential biological markers for predicting the tendency of CBD malignancy or diagnosing the occurrence of CBD malignancy.