TTYH3 expression and Prognostic values in HCC
The mRNA levels of TTYH3 in HCC were obtained from UALCAN database. As shown in Fig. 1A, mRNA expression of TTYH3 was found to be significantly up-regulated in HCC tissues compared to normal samples (p < 0.01), Then We used UALCAN to analyze the prognostic values of the mRNA expression of TTYH3 in HCC patients. As was shown in Fig. 1B, mRNA expression in members of the TTYH3 was significantly associated with prognosis in patients with HCC disease. We found that higher mRNA expression of TTYH3 was significantly associated with shorter OS of HCC patients, then we used GEPIA to analyze the prognostic value of the expression of TTYH3 in HCC patients, The results showed that the expression level of TTYH3 was significantly correlated with prognosis in HCC with poor OS and DFS, These results indicate that TTYH3 mRNA was significantly associated with prognosis in patients with HCC and can be used as a biomarker to predict survival in patients with HCC.
Relationship between TTYH3 and clinical pathological in HCC patients
we analyzed the relationship between TTYH3 with clinicopathological parameters of HCC patients by UALCAN and GEPIA, As was shown in Fig. 2, The TTYH3 was correlated with individual cancer stage, indicating that patients with more advanced cancer stages tended to have higher TTYH3 expression. the results above suggested that TTYH3 was significantly associated with clinicopathological parameters in HCC patients.
Genetic mutation and Prognostic values of TTYH3 in HCC patients
We used cBioPortal to analyze the genetic mutation of TTYH3 in HCC patients. Based on Fig. 3, the mutation rate of TTTYH3 was 6% in 366 samples. What’s more, the association between genetic mutation and the prognosis of HCC patients was explored. And a statistically significant correlation was found between genetic mutation of TTYH3 and OS (p = 0.0353) in HCC patients, and DS was also statistically significant (p = 0.0128), However, there was no statistical difference with DF.
Methylation analysis of TTYH3 in HCC
In order to find the main causes of aberrant expression of TTYH3 in HCC, we used UACLAN database as well as MEXPRESS database. As was shown in Fig. 4, We found that TTYH3 is hypomethylation in HCC by UACLAN database (p < 0.01), Then, the methylation level of TTYH3 detected by MEXPRESS database. The result revealed that the promoter region of TTYH3 showed significantly lower methylation levels (p < 0.05) in HCC compared to normal tissues, the result showed the negative correlation between TTYH3 expression and promoter methylation.
TTYH3 positively correlates with immune cell infiltration in HCC
the correlation of TTYH3 expression level with immune cell infiltration level was evaluated. As presented in Figs. 5A, TTYH3 expression was significantly positively associated with all analyzed immune cells, including B cell, CD8 + T cell, CD4 + T cell, macrophage, neutrophil, and dendritic cell in HCC, As were shown in Fig. 5B-C and Fig. 6, the TISIDB database was used to infer the relations between abundance of TILs and expression of TTYH3. The relations between abundance of 28 TIL types and expression of TTYH3 was weakly to moderately correlated. Specifically, TTYH3 expression was positively closely related with infiltrating levels of Act CD4 (r = 0.222, P = 1.6e-05) and Act DC (r = 0.257, P = 5.58e-07), then we used GEPIA to analyze the correlation between TTYH3 and monocytes, TAM and macrophage-related genes and markers (Fig. 7). Moreover, the correlation between TTYH3 expression and the marker genes of immune cells implicate the role of TTYH3 in regulating tumor immunology. First, gene markers of M1 macrophages such as PTGS2 and IRF5 showed positive correlations with TTYH3 expression, M2 macrophage markers such as CD163, VSIG4, and MS4A4A also showed moderate and strong correlations.
Networks Analyses and Functional Enrichment Analysis of TTYH3 and their Neighboring Genes in HCC patients
similar genes of the TTYH3 (30 in total) obtained from GEPIA and Metascape were used for GO enrichment to explore the interaction between similar genes. Based on 30 adjacent genes, the online tools of Metascape and STRING were used for functional and pathway enrichment analysis, and a PPI interaction network was established to explore the biological classification of TTYH3. The functions of TTYH3 neighboring genes were predicted by analyzing GO and KEGG in Metascape. The GO enrichment items were classified into four functional groups,KEGG pathway, biological process group, molecular function group, and cellular component group (Figs. 8). The TTYH3 and similar genes was Enriched in the following information, bone morphogenesis, regulation of proteolysis, regulation of anatomical structure size, carbohydrate metabolic process, immune response-activating cell surface receptor signaling pathway, actin cytoskeleton organization, angiogenesis (BP); and focal adhesion, polymeric cytoskeletal fiber (CC); and glycosaminoglycan biosynthesis, Tuberculosis in KEGG pathway. The PPI network interaction of TTYH3 was conducted by String to seek possible downstream targets and mechanism research, and it was found that BEST1、BEST2、BEST3、BEST4、ZNF467、CLCC1、GLRB、ANO1、IQCE、ANO2 and other genes could be used as the target genes for further research and analysis.