Fig. 7

circTOLLIP stimulates EMT by miR-516a-5p/PBX3 axis. a Clustering heatmap showing the RNA-seq profiles in vector and PBX3-overexpressing 97H cells. b-c Gene set enrichment analysis (GSEA) of genes from RNA-seq profiles and TCGA database comparing with EMT_PATHWAY. d Among the EMT-related genes in TCGA, those with the highest positive correlation with PBX3 expression were selected and intersected with the differentially upregulated genes in the RNA-seq profiles. The top four genes (PTX3, COL6A2, CCN2, and MATN3) were verified to be upregulated in 97H cells with PBX3 overexpression compared with vector cells. And these genes were also upregulated in HLF and 97H cells with circTOLLIP overexpression (e, f) though qRT–PCR analysis. g Western blot analysis of PBX3 protein expression in circTOLLIP-overexpressing cells with or without transfection of miR-516a-5p mimic or PBX3 siRNA. h-i CCK-8 and colony formation assays to evaluate the cell proliferation ability respectively in four group cells. j-l Scratch wound healing assay and cell migration and invasion assays to evaluate the cell metastatic ability