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Fig. 4 | BMC Cancer

Fig. 4

From: Cellular retinol binding protein-1 inhibits cancer stemness via upregulating WIF1 to suppress Wnt/β-catenin pathway in hepatocellular carcinoma

Fig. 4

CRBP-1 inhibits Wnt/β-catenin signaling pathway by upregulating WIF1. (A) Heatmap of RNA-seq analysis for differentially expressed cancer stemness related genes (fold change ≥1.5, p < 0.05) in response to CRBP-1 overexpression. (B) Gene set enrichment analysis (GSEA) was performed to investigate the activation of Wnt/β-catenin signaling pathway against CRBP-1 expression in HCC tissue samples. Dataset were obtained from TCGA (n = 374). (C) Correlation analysis between CRBP-1 and WIF1 in HCC tissue samples was performed with Spearman correlation analysis. Dataset were obtained from TCGA (n = 374). (D) qPCR analysis of the expression of WIF1 and β-catenin (CTNNB1) in CRBP-1 overexpressed PLC/PRF/5 cells and control cells. Data shown represent the mean ± SD from three independent experiments. (E) Western-blot analysis of the expression of WIF1, β-catenin, and phosphorylated β-catenin in CRBP-1 overexpressed PLC/PRF/5 and Huh7 cells as well as control cells. (F) Western-blot analysis of the expression of β-catenin in CRBP-1 overexpressed PLC/PRF/5 cells and control cells, which were treated with 10 μM of MG-132 under indicated time. (G) Western-blot analysis of the expression of c-Myc and Cyclin D1 in CRBP-1 overexpressed PLC/PRF/5 and Huh7 cells as well as control cells. (H) qPCR analysis of the expression of c-Myc and Cyclin D1 in CRBP-1 overexpressed PLC/PRF/5 and Huh7 cells as well as control cells. Data shown represent the mean ± SD from three independent experiments. (I) The transcriptional activity analysis of β-catenin in CRBP-1 overexpressed cells and control cells by using TOP/FOP-Flash luciferase reporter system. Data shown represent the mean ± SD from three independent experiments

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