| Objective: Exploring the new pyroptosis-related subtypes and prognostic markers to improve survival rate of gastric cancer(GC).Methods: The clinical and transcriptome data of GC patients were downloaded from TCGA(n=407)and GEO(n=433)database.The expression of pyroptosis-related genes and "Consensus Cluster Plus" package were used to cluster the GC patients."Survminer" package was used for survival analysis and the single-sample gene set enrichmentanalysis(ss GSEA)algorithm was used to compare differences in immune cell infiltration between two different subtypes.Lasso regression and Cox regression were used to construct the prognostic risk model.The gene involved in model construction was filtered by using clinical correlation analysis.The differences in the expression levels of APOD,TGM2 and SLC2A3 in paracancellular tissue and tumor tissue and their effects on the prognosis of gastric cancer were analyzed using a public database.The KEGG analyses were performed by applying the GSEA software.The CIBERSORT algorithm was used to evaluate immune cell content and explore its correlation with target genes.The online sites were used to explore the association between target genes and immunoinfluential factors and drug sensitivity.The public databases were used to explore the correlation between the expression level of target genes and pyroptosis-related genes and pyroptosis-related subtypes.Finally,immunohistochemistry and q T-PCR were used to verify the results of bioinformatics analysis.Results: There were significant differences in the prognostic level between two pyroptosis-related subtypes.The content of 22 kinds of immune cells in high prognosis type was higher than those in low prognosis type(p<0.01).There were statistically significant differences in different age groups(p<0.01)and N stages(p=0.04)between the two pyroptosis-related subtypes.The results of public database analysis showed that the expression level of APOD in GC tumor tissue was lower than that in paracancer tissue,while the expression level of TGM2 and SLC2A3 in GC tumor tissue was higher than that in paracancer tissue.Survival analysis showed that high expression of APOD,TGM2 and SLC2A3 was not conducive to GC prognosis.Clinical correlation analysis showed that the expression levels of APOD and SLC2A3 were statistically significant in different T stages.The GSEA results showed that high expression of APOD group was enriched in cell adhesion,leukocyte transendothelial migration,and gap junction pathway.The high-expression group of TGM2 was enriched in chemokine signaling,regulation of actin cytoskeleton cytokine and cytokine receptor interaction pathways,while the high-expression group of SLC2A3 was enriched in cell adhesion molecules,pathways in cancer,neurotrophin signaling pathway.The results of immune infiltration showed that the contents of follicle-assisted T cells,CD4+ memory-activated T cells,resting NK cells and neutrophils were lower in APOD high expression group.The contents of CD4+memory resting cells was lower in TGM2 high expression group.The contents of CD4+ memory resting cells and regulatory cells were lower in SLC2A3 high expression group.APOD was positively correlated with cell cycle detection point kinase inhibitor AZD-7762,multiple kinase inhibitor KW-2449 and JAK2 inhibitor TG-101348(0<rho<0.3,P<0.05).TGM2 was positively correlated with Dasatinib,CXCR2 antagonist SB225002,carbic anhydrase inhibitor indisulam and multikinase inhibitor rigosetib(0<r<0.4,P< 0.05).Immunohistochemical and qt-PCR results showed that APOD was lower in tumor tissue,while TGM2 and SLC2A3 were higher in tumor tissue.Moreover,high expression of APOD and SLC2A3 has poor prognosis for GC patients.Conlusion: The prognosis of GC patients could be well evaluated according to pyroptosis-related subtypes,and APOD,TGM2 and SLC2A3 have the potential to be new prognostic markers and therapeutic targets for GC. |
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