The Mechanism Of GNB1 Promoting Proliferation, Invasion And Metastasis Of Hepatoblastoma And Multi-omics Studies Reveal The Role Of Macrophage-endothelial Cell Interaction In The Tumor Microenvironment Of Hepatoblastom

Mechanism of GNB1 promoting proliferation,invasion and metastasis of hepatocellular carcinomaPurposeHepatoblastoma(HB)mainly occurs in patients at an early age and is the most common primary malignancy.Today,the incidence of hepatoblastoma worldwide is increasing year by year at an incidence of about 3% in people under 18 years of age.In recent years,despite the increasing diversity of diagnosis and treatment methods for hepatoblastoma and the increasing concept of multidisciplinary treatment,the overall survival rate(Overall Survival,OS)of children with hepatoblastoma has been well improved,but the treatment of advanced and refractory hepatoblastoma remains a huge clinical problem.It is therefore important to find early indicators for prognosis prediction in hepatoblastoma.In this study,we identify differentially expressed genes between hepatoblastoma and adjacent normal tissues,explore the relationship between GNB1 expression in clinical hepatoblastoma specimens and related clinical parameters,and analyze and identify the role of GNB1 in hepatoblastoma using cytofunctional experiments.The aim is to explore the pathogenesis of hepatoblastoma and to find potential therapeutic targets.methodIn this study,we analyzed the expression of GNB1 in hepatoblastoma by bioinformatics analysis.In order to understand the role of GNB1 in the progression of hepatoblastoma,we collected clinical tissue samples from pediatric surgery department of the Affiliated Hospital of Qingdao University and double-verified the differential expression of GNB1 in hepatoblastoma and adjacent non-cancerous tissues using real-time quantitative fluorescence PCR assay and Western blot assay.Further investigate the effect of GNB1 on the function of hepatoblastoma cells,lentiviral transfection of hepatoblastoma cell line Hep G2 cells resulted in overexpression of GNB1,and the expression of GNB1 in Hep G2 was knocked down by transient transfection technique.Transwell assay and scratch assay were performed in Hep G2 cell line to verify the effect of GNB1 on hepatoblastoma cell migration and invasion;CCK8 assay was performed on Hep G2 cells to verify the effect of GNB1 on hepatoblastoma proliferation.Finally,the mechanism of GNB1 on Wnt/β-Catenin signaling pathway was explored by WB assay.Results1.The database explores differentially expressed genes between hepatoblastoma tissues and adjacent adjacent normal tissues,screens and identifies key genes,and constructs a PPI interaction network.2.Enrichment analysis for differential genes,3.Bioinformatic analysis of the target gene GNB1 showed that GNB1 was highly expressed in a variety of malignancies,whereas in hepatic malignancies,GNB1 expression was upregulated relative to adjacent non-cancerous tissues.However,survival analysis was performed for G NB1,and the overall survival rate was significantly lower in the high GNB1 expression group than in the low expression group.4.Hepatoblastoma clinical tissue samples RNA was extracted,reverse transcribed and a c DNA library was constructed,and real-time quantitative fluorescence PCR experiments showed that the GNB1 gene was up-regulated in hepatoblastoma,consistent with database analysis results.Further extraction of clinical hepatoblastoma tissue samples and Western blot experiments showed that the GNB1 gene was up-regulated in hepatoblastoma.5.GNB1 expression was identified in the hepatoblastoma cell line Hep G2,and GNB1 was overexpressed and knocked down by constructing lentivirus and transient transfection techniques,respectively.Further cell functional experiments were performed on GNB1,and CCK8 assay revealed that the proliferation ability of hepatoblastoma cells was significantly enhanced after overexpression of GNB1 in the Hep G2 line,and knockdown of GNB1 expression was significantly reduced.Transwell assay and cell scratch assay verified that invasion and migration of hepatoblastoma cells were enhanced after overexpression of GNB1 in Hep G2 cells,and knockdown of GNB1 expression significantly decreased.6.Western blot experiments showed that GNB1 plays an important role in regulating the EMT mechanism of hepatoblastoma through Wnt/β-catenin signaling pathway.ConclusionGNB1 is up-regulated in hepatoblastoma tissue,enhancing the ability of GNB1 to regulate Wnt/β-catenin signaling pathway in hepatoblastoma and may serve as a prognostic marker for hepatoblastoma and may be a potential therapeutic target for hepatoblastoma.Multiomics reveals the role of macrophage-vascular endothelial cell interactions in the hepatoblastoma tumor microenvironmentPurpose: Hepatoblastoma is a common tumor in childhood,characterized by immature histology and diverse cell lineages.The purpose of this study is to identify the genes that are abnormally expressed in hepatoblastoma,and to explore and verify the targets of intercellular communication that affect the tumor immune microenvironment.Methods: Through comprehensive analysis of gene expression from GSE133039 and GSE180664 data sets,the differentially expressed genes in cancer tissues and adjacent tissues were obtained.GO and KEGG enrichment analysis is used to predict the biological function and signal transduction pathway of differential expression gene enrichment.Use cytoscape to build PPI network to filter hubgene;Construct correlation analysis of immune cell infiltration to infer the correlation between immune cells.Combined with single-cell transcriptome data,further reveal the relationship between cells and signal targets of cell communication.Results: 58 differentially expressed genes with high expression and 94 differentially expressed genes with low expression were obtained from the two data sets of hepatoblastoma.They were mainly involved in the signal transduction related to metastasis.PPI network screened 50 hubgenes.The correlation analysis of immune cell infiltration of different genes showed that macrophages were significantly correlated with endothelial cells.Combined with the analysis of single-cell transcriptome data,hepatoblastoma was divided into 11 cell subpopulations,and 16 genes in hubgene were expressed in different cell subpopulations,in which LFNG was highly expressed in macrophages and monocytes,which served as the target of intercellular communication to promote the development of hepatoblastoma.Conclusion: In this study,we identified the genes that were abnormally expressed in hepatoblastoma,and affected the cellular communication of hepatoblastoma through LFNG target,thus affecting the progress of tumor.Therefore,LFNG may become a therapeutic target for hepatoblastoma.