Molecular Mechanism Of GPSM2 Regulating HCC And Potential Traditional Chinese Medicine Based On Bioinformatic

Objective:1.To explore the potential regulatory mechanism of G protein signaling modulator2(GPSM2)in hepatocellular carcinoma(HCC)based on bioinformatics approach.2.To screen key gene targets based on the interaction between GPSM2 related genes and reverse mining potential drugs and active ingredientsMethods:1.The expression of GPSM2 in 33 tumor tissues including HCC was analyzed using TCGA database and GEO database;Kaplan-Meier plotter database was used to analyze the relationship between the expression level of GPSM2 and the survival of HCC patients,and the relationship between GPSM2 and clinical parameters of patients was analyzed using TCGA HCC data and clinical data in cell lines and clinical tissue samples to validate the expression of GPSM2;online analysis tools to analyze the reciprocal proteins of GPSM2 and the biological processes involved in regulation;R package to perform GO/KEGG and GSEA functional enrichment analysis of GPSM2;application of TIMER,TISDB database to analyze the relationship between GPSM2 expression and HCC immune cell infiltration and immune subtypes and analyze the relationship between GPSM2 and immune checkpoint-related genes;analyze the mutation and methylation of GPSM2 in HCC using c Bio Portal,SMART App online tool;predict and analyze the non-coding RNA upstream of GPSM2based on star Base3.0 database.2.Using STRING database,the Cytoscape software to obtain key genes in GPSM2-related gene interactions regulatory network,GEPIA2 database to assess the expression differences and prognostic impact of key genes in HCC,and the correlation between key genes and GPSM2;using Coremine MEDICAL database to reverse predict the high frequency Chinese medicine acting on key genes,in TCMSP database To obtain the corresponding information of active ingredients and targets of traditional Chinese medicine.Results:1.The GPSM2 gene was abnormally highly expressed in a variety of tumors including HCC(P<0.05),and the expression level of GPSM2 was associated with the pathological stage,T-stage,tumor histological grade,tumor recurrence status during follow-up,and prothrombin time of HCC patients;the results of survival analysis showed that high GPSM2 gene expression was associated with poor prognosis and patients’survival time was significantly reduced.We also experimentally verified that the m RNA and protein levels of GPSM2 were expressed higher in HCC cell lines than in normal hepatocyte lines,and the results of immunohistochemical staining showed that the expression level of GPSM2 was higher in HCC tissues than in paraneoplastic tissues,and was associated with the clinicopathological stage of patients.2.Functional enrichment and other analyses showed that in HCC,GPSM2 may regulate biological processes such as cell mitotic cycle,DNA repair,and immune microenvironment,and GPSM2 is closely associated with PI3K/AKT,MAPK,Wnt,Notch and other pro-oncogenic signals.3.By further correlation analysis,GPSM2 expression was significantly and positively correlated with DNA repair-related genes,with the infiltration levels of CD4~+T cells,CD8~+T cells,macrophages,neutrophils and dendritic cells in HCC,and with the expression of immune checkpoint-related genes in HCC.4.Combined multi-omics analysis showed that the aberrant high expression of GPSM2 in HCC was associated with the effects of methylation and copy number variation,LINC00858/hsa-mi R-22-3p,MIR4435-2HG/hsa-mi R-22-3p and BX284668.6/hsa-mi R-22-3p as non-coding RNA regulatory pathways upstream of GPSM2.5.Five key genes,CDK1,CCNB1,CCNA2,KIF11,and ASPM,were screened from the reciprocal network of GPSM2-associated genes,and high expression of these five genes in HCC were negatively correlated with prognosis and highly positively correlated with GPSM2;by inverse prediction of potential herbal medicines,we obtained nine herbal medicines and their active ingredients with regulatory effects on key genes,namely,scutellaria barbata,rhubarb,burnet,dried ginger,citrus aurantium,magnolia officinalis,ginger,strychnos,evodia rutaecarpa.Conclusion:1.GPSM2 expression was significantly upregulated in HCC and was closely associated with poor patient prognosis and tumor immune cell infiltration;aberrant GPSM2expression was regulated by DNA methylation,copy number variation,and non-coding RNA;GPSM2 is involved in the regulation of the mitotic cycle of cells multiple signaling pathways contribute to the development of HCC.2.Initially,we found that nine traditional Chinese medicines and active ingredients,namely,scutellaria barbata,rhubarb,burnet,dried ginger,citrus aurantium,magnolia officinalis,ginger,strychnos,evodia rutaecarpa,had regulatory effects on key genes in the potential regulatory network of GPSM2,which provided a preliminary basis for further research on traditional Chinese medicines for the treatment of HCC.