Fig 2: Significant TICRR and PPIF-related genes and pathways in EC obtained by GSEA and GSVA. (A) The most involved pathways in high-expression group of TICRR obtained by GSEA analysis. (B) Transcriptional expression profiles of the significant genes were shown in a heat map. (C) The most common functional gene sets in high-expression group of PPIF by GSEA. (D) Transcriptional expression profiles of the significant genes related with PPIF were shown in a heat map. (E, F) GSVA-derived clustering heatmaps of differential pathways for TICRR and PPIF, respectively.

Fig 3: Validation of hub genes by cell experiments in vitro. (A) Identification of differentially expressed genes by qPCR assay in both normal endometrial epithelial cells and endometrial cancer cell lines. (B) Western blot assay showing the expression of TICRR and PPIF in EC cell lines and normal endometrial epithelial cells. (C) The effect of different siRNA on TICRR and PPIF gene silence by western blotting respectively. (D) TICRR and PPIF expression were examined by RT-PCR after transfected with siRNA for 24h. (E) Relevant molecular targets were verified by qRCR after transfection of siTICRR and siPPIF for 24hrs respectively. (F) Cell growth and clone formation of siTICRR transfected cells with or without MPA. (G) Cell growth and clone formation of siPPIF transfected cells with or without MPA. Data were shown as mean ± SD; *p < 0.05; **p < 0.01; ***p < 0.001.

Fig 4: IHC staining and functional verification of TICRR and PPIF genes. (A), The expression of TICRR and PPIF in endometrial cancer and paracancer samples were assessed using IHC staining and the results of statistical analysis. (B) Analysis of apoptosis by Flow cytometry after transfection with siTICRR and siPPIF respectively. (C) Hec-1B-siTICRR cells were subjected to migration and invasion assays in the presence or absence of MPA. (D) Migration and invasion ability of siPPIF transfected cells in the presence or absence of MPA. (E) Wound-healing assays for Hec-1B-siTICRR cells. (F) Wound-healing assays for Hec-1B-siPPIF cells. Data were shown as mean ± SD; *p < 0.05; **p < 0.01; ***p < 0.001.