| Objective: 1.To explore the expression of sperm associated antigen 5(SPAG5)gene in liver cancer and its relationship with prognosis and immune infiltration of patients with liver cancer by bioinformatics analysis,and to predict the possible molecular mechanism of SPAG5 in the regulation of liver cancer,so as to provide a reference for further research of the value of SPAG5 in the diagnosis and treatment of liver cancer.2.The main molecular targets and signal pathways of curcumin acting on liver cancer were analyzed by network pharmacology,and the effect of curcumin on PI3 K / NF-k B pathway in Hep G2 cells was preliminarily verified by RT-q PCR and Western Blot.Methods: 1.Analyze the expression of SPAG5 gene in liver cancer patients by Oncomine and TIMER database;analyze the expression difference of SPAG5 gene in different clinical stages of liver cancer patients by GEPIA database;analyze the relationship between SPAG5 gene expression and prognosis of liver cancer patients by Kaplan-Meier Plotter database;analyze the relationship between SPAG5 gene expression and immune infiltration of liver cancer by TIMER and GEPIA database;TCGA database and GSEA software were used to analyze the molecular mechanism of SPAG5 gene acting on liver cancer.2.Use Pub Chem,Pharmmapper,Uniprot,Genecard,String database,and Cytoscape software to obtain the main molecular targets of curcumin acting on liver cancer and draw a molecular interaction network diagram;use cluster Profiler package in R software for the main targets of curcumin acting on liver cancer Enrichment analysis of GO and KEGG was performed;RT-q PCR and Western Blot were used to detect PI3 K and NF-k B m RNA and protein expression levels of curcumin-treated liver cancer Hep G2 cells.Results: 1.Oncomine database and TIMER database showed that SPAG5 gene was highly expressed in liver cancer(P < 0.01);GEPIA database showed that the level of SPAG5 in patients with stage II and stage III liver cancer was significantly higher than that in patients with stage I liver cancer(P < 0.05);Kaplan-Meier Plotter database showed that SPAG5 gene high expression and overall survival time(OS)ofliver cancer patients,Progression-free survival time(PFS),relapse-free survival time(RFS),and disease-specific survival time(DSS)were significantly related to shortening(P < 0.01);TIMER and GEPIA database analysis found that SPAG5 gene expression was significantly correlated with the infiltration level of CD8 + T cells,macrophages,neutrophils,B cells,dendritic cells and T-depleted cells in liver cancer(P < 0.01);GSEA analysis showed that SPAG5 mainly affected liver cancer by regulating cell cycle,DNA replication,autophagy,T cell receptor,toll like receptor,p53,m TOR,Notch,Wnt and other signaling pathways(FDR < 0.05,NP < 0.05).2.Network pharmacology analysis found that curcumin has 66 main targets for liver cancer.GO functional enrichment analysis found that curcumin mainly affects transcription factor activation,protein kinase activation,receptor binding,receptor activation,and five aspects of biological binding,KEGG pathway enrichment analysis found that curcumin mainly affected PI3K/AKT signaling pathway,MAPK signaling pathway,tumor proteoglycan signaling pathway,tumor central carbon metabolism signaling pathway,etc.RT-q PCR and Western Blot test results it indicates that curcumin can inhibit the m RNA and protein expression levels of PI3 K and NF-k B.Conclusions: 1.Bioinformatics analysis shows that SPAG5 gene is highly expressed in HCC,and the high expression of SPAG5 is related to the immune infiltration and poor prognosis of HCC patients.SPAG5 can promote the proliferation,migration,invasion,inhibition of apoptosis,and regulate tumor immunity by regulating multiple signal pathways.2.Network pharmacology analysis showed that curcumin could intervene the development of liver cancer through multiple targets and signal pathways.Experiments verify showed that curcumin could inhibit the PI3K/NF-k B pathway in Hep G2 cells. |
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