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Junxue Tu,1 Jingjing Chen,2 Meimei He,1 Hongfei Tong,3 Haibin Liu,3 Bin Zhou,3 Yi Liao,3 Zhaohong Wang31Department of Pharmacy, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, People’s Republic of China; 2Department of Breast Surgery, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, People’s Republic of China; 3Department of Hepatobiliary Surgery, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, People’s Republic of ChinaBackground: Hepatocellular carcinoma (HCC) is the second leading cause of death among cancers worldwide. In this study, we aimed to identify the molecular target genes and detect the key mechanisms of HCC. Three gene expression profiles (GSE84006, GSE14323, GSE14811) and two miRNA expression profiles (GSE40744, GSE36915) were analyzed to determine the molecular target genes, microRNAs (miRNAs) and the potential molecular mechanisms in HCC.Methods: All profiles were extracted from the Gene Expression Omnibus database. The identification of the differentially expressed genes (DEGs) was analyzed by the GEO2R method. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and gene ontology (GO) enrichment analysis performed database for Integrated Discovery, Visualization and Annotation. The miRNA-gene network and protein–protein interaction (PPI) network were correlated by the Cytoscape software. The key target genes were identified by the CytoHubba plugin, Molecular Complex Detection (MCODE) plugin and miRNA-gene network. The identified hub genes were testified for survival curve using the Kaplan–Meier plotter database.Results: Expression profiles had 592 overlapped DEGs. The majority of the DEGs were enriched in membrane-bounded organelles and intracellular membrane-bounded organelles. These DEGs were significantly enriched in metabolic, protein processing in the endoplasmic reticulum and thyroid cancer pathways. PPI network analysis showed these genes were mostly involved in the pathogenic Escherichia coli infection and the regulation of actin cytoskeleton pathways. Combining these results, we identified 10 key genes involving in the progression of HCC. Finally, PLK1, PRCC, PRPF4 and PSMA7 exhibited higher expression levels in HCC patients with poor prognosis than those for lower expression via Kaplan–Meier plotter database.Conclusion: PLK1, PRCC, PRPF4 and PSMA7 could be potential biomarkers or therapeutic targets for HCC. Meanwhile, the metabolic pathway, protein processing in the endoplasmic reticulum and the thyroid cancer pathway may play vital roles in the progression of HCC.Keywords: hepatocellular carcinoma, DEGs, bioinformatic analysis, KEGG pathway, PPI network, miRNA-gene network

During Toll-like receptor 4 signaling, the Toll/interleukin-1 receptor domain is essential for interactions with downstream Toll/interleukin-1 receptor domain-containing adaptor proteins. The aim of this study is to investigate the role of the Toll/interleukin-1 receptor domain in the Toll-like receptor 4 signaling pathway during hepatic ischemia and reperfusion injury.We genetically blocked the function of Toll/interleukin-1 receptor domain in mice and examined the effect on Toll-like receptor 4 signaling and the response to hepatic ischemia and reperfusion.University research laboratory.Male BALB/c mice.Male BALB/c mice were hydrodynamically administrated the target gene plasmid pTIR-IRES2-EGFP, empty vector containing enhanced green fluorescent protein, or normal saline. Animals underwent 90 minutes of partial hepatic ischemia, followed by 6 or 24 hours of reperfusion. Hepatic injury was assessed by measuring serum alanine transaminase, hepatic histology, and malondialdehyde. The expression of inflammatory cytokines and nuclear factor-κB phosphorylation was examined in liver tissues. Hepatic apoptosis was evaluated by caspase-3 assays, terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick-end labeling staining, and the activation of Jun N-terminal kinase and p38 mitogen-activated protein kinase.Blocking the Toll/interleukin-1 receptor domain resulted in markedly lower serum alanine transaminase levels, reduced histologic injury, and lower malondialdehyde levels following 6 or 24 hours of hepatic reperfusion than mice receiving the vector alone or normal saline. Anti-Toll/interleukin-1 receptor treatment also reduced Toll-like receptor 4 expression and disrupted Toll/interleukin-1 receptor-Toll/interleukin-1 receptor interactions in Toll-like receptor 4 signaling pathways. Blocking the Toll/interleukin-1 receptor domain also prevented Toll-like receptor 4-mediated mitogen-activated protein kinase activation (via Jun N-terminal kinase and p38 mitogen-activated protein kinase), an activation that mediated liver ischemia and reperfusion injury via caspase-3 activation, resulting in increased hepatocellular apoptosis. Lastly, blocking the Toll/interleukin-1 receptor domain decreased inflammatory cytokine production by inhibiting nuclear factor-κB activation.Toll/interleukin-1 receptor domain inhibition disrupts the interaction of Toll-like receptor 4 with its adaptor proteins, which abrogates downstream signaling pathways and prevents the activation of nuclear factor-κB and Jun N-terminal kinase/p38. This reduction in signaling consequently reduces hepatic inflammation, cell apoptosis, and hepatic damage. Toll/interleukin-1 receptor domain-targeted therapy thus represents a new approach to inhibit the intracellular Toll-like receptor 4 signaling pathway and reveals novel therapeutic target sites, which will facilitate the development of specific therapeutic agents.