Journal: Molecular Cancer

Article Title: Upregulation of microRNA-122 by farnesoid X receptor suppresses the growth of hepatocellular carcinoma cells

doi: 10.1186/s12943-015-0427-9

Figure Lengend Snippet: The level of FXR is positively correlated with that of miR-122. a and b The expression of FXR mRNA ( a ) and mature miR-122 ( b ) in 20 human HCC tissues and the corresponding adjacent noncancerous tissues was detected by qRT-PCR. c The correlation between the levels of FXR and miR-122 in HCC tissues was analyzed using Pearson’s test ( R 2 = 0.61, P < 0.01). d The expression of FXR mRNA and mature miR-122 in HCC cell lines (HepG2, Hep3B, Huh7, PLC, SMMC-7721, MHCC97L and MHCC97H) and hepatic cell line L02 was assayed by qRT-PCR. e The correlation between the levels of FXR and miR-122 in HCC cell lines was analyzed using Pearson’s test ( R 2 = 0.95, P < 0.01). β-actin was used as a control for FXR examination, while U6 snRNA as a control for miR-122 detection

Article Snippet: The other HCC cell lines including Huh7, PLC, SMMC-7721, MHCC97L and MHCC97H, and hepatic cell line L02 were purchased from China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Signal Transduction and Targeted Therapy

Article Title: Dysadherin/YAP axis fuels stem plasticity and immune escape in liver cancer

doi: 10.1038/s41392-025-02520-4

Figure Lengend Snippet: Dysadherin promotes cancer stem-like cell features and tumor progression in HCC. a UMAP plot of single-cell transcriptomes from HCC patient samples ( GSE166635 ; n = 2). Cell lineage trajectories were inferred using Monocle2, highlighting the progression toward a malignant state. The expression of CSC- and malignancy-associated genes was mapped along the trajectory. b UMAP and GSEA of tumor clusters from GSE166635 , comparing dysadherin high ( n = 1647) and dysadherin low ( n = 2853) tumor cells. c GSEA of DEGs between dysadherin high and dysadherin low tumors from bulk RNA-seq data ( GSE9843 ; FDR < 0.05). d Violin plots showing FXYD5 expression levels in normal liver tissue ( n = 50) and primary HCC tissues ( n = 371; TCGA-LIHC), and stratified by tumor grades. e Kaplan–Meier survival analysis for overall survival based on FXYD5 expression in TCGA-LIHC cohort. Statistical significance was assessed by log-rank tests. f Limiting dilution sphere formation assay (left) and Aldefluor assay (right) evaluating the effect of dysadherin expression on CSC properties. Scale bar = 50 μm. g Heatmaps of CSC- and malignancy-related gene expression in HCC cells with dysadherin overexpression or knockdown. h In vivo limiting dilution assay (LDA) assessing the tumor-initiating potential of SK-Hep1 cells with or without dysadherin-knockdown (DYS KD ). i Tumor growth curves from the in vivo limiting dilution assay comparing tumor-initiating capacity of DYS KD versus control (WT) SK-Hep1 cells across multiple inoculation doses (0.5–10 × 10 3 cells/mouse). j Heatmaps showing the expression of CSC-associated genes in dysadherin KD versus control SK-Hep1 cells. k Schematic view of DEN/CCl 4 -induced HCC mouse model. l Representative gross liver images and immunofluorescence staining of liver sections from Fxyd5 +/+ and Fxyd5 −/− mice. Scale bar = 200 μm. m Expression of HCC markers in tumor (T) and paired adjacent normal tissues (P) from DEN/CCl 4 -treated mice. n Quantification of tumor number and distribution of tumor sizes, including largest tumor per mouse. o Tumor incidence in mice at the indicated time points. p Heatmap showing expression levels of CSC- and malignancy-associated genes in liver tissues from mice. Data are presented as means ± SEM. Statistical significance was determined by unpaired two-tailed Student’s t-tests and one-way ANOVA with Dunnett’s multiple comparison tests. *, ** and *** indicate p < 0.05, p < 0.01, and p < 0.001

Article Snippet: The human HCC cell lines, HepG2, SNU368, and SK-Hep1 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Republic of Korea).

Techniques: Expressing, RNA Sequencing, Tube Formation Assay, Gene Expression, Over Expression, Knockdown, In Vivo, Limiting Dilution Assay, Control, Immunofluorescence, Staining, Two Tailed Test, Comparison

Journal: Signal Transduction and Targeted Therapy

Article Title: Dysadherin/YAP axis fuels stem plasticity and immune escape in liver cancer

doi: 10.1038/s41392-025-02520-4

Figure Lengend Snippet: Dysadherin acts as an upstream regulator of YAP signaling to promote malignancy in HCC. a GSEA of DEGs between dysadherin high and dysadherin low HCC tumors ( GSE9843 ), highlighting enrichment of YAP-related oncogenic signatures. b Left: UMAP plot of tumor cell clusters from GSE166635 , color-coded by FXYD5 expression. Right: UMAPs and violin plots showing YAP signature scores and expression of YAP target genes ( CTGF , CYR61 ) in FXYD5 high versus FXYD5 low clusters. c Kaplan–Meier survival analysis of overall survival in TCGA-LIHC cohort stratified by high or low expression of FXYD5 and YAP target genes ( CTGF or CYR61 ). Significance was assessed by log-rank test. d Immunofluorescence (IF) staining of dysadherin and nuclear YAP in paired normal and tumor tissues from HCC patients ( n = 14). Representative images and quantification of nuclear YAP signal intensity are shown. Scale bar = 100 μm. e Violin plots comparing dysadherin expression and nuclear YAP intensity across histological grades of HCC in patient tissues ( n = 14). f RT-qPCR and immunoblot analyses of dysadherin, total YAP, and active YAP expression in normal hepatic (THLE-3) and HCC cell lines. g Heatmaps and immunofluorescence staining showing expression of YAP target genes and subcellular localization of active YAP in dysadherin-overexpressing (OE) and knockdown (KD) cells. Scale bar = 10 μm. h Immunoblotting analysis of active YAP (non-phosphorylated) and phospho-YAP (S127) following dysadherin OE or KD. i Heatmap showing YAP target gene expression in tumor from DEN/CCl₄-treated Fxyd5 +/+ and Fxyd5 −/− mice. j Immunoblotting of YAP pathway components and downstream targets (CTGF) in liver lysates from Fxyd5 +/+ and Fxyd5 −/− mice. Data are presented as means ± SEM. Statistical significance was determined by unpaired two-tailed Student’s t-tests and one-way ANOVA with Dunnett’s multiple comparison tests. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The human HCC cell lines, HepG2, SNU368, and SK-Hep1 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Republic of Korea).

Techniques: Expressing, Immunofluorescence, Staining, Quantitative RT-PCR, Western Blot, Knockdown, Targeted Gene Expression, Two Tailed Test, Comparison

Journal: Signal Transduction and Targeted Therapy

Article Title: Dysadherin/YAP axis fuels stem plasticity and immune escape in liver cancer

doi: 10.1038/s41392-025-02520-4

Figure Lengend Snippet: Dysadherin promotes CSC features through YAP activation in HCC. a Upstream regulator analysis using dysadherin-correlated transcriptional signatures in HCC tumors ( GSE9843 ), predicting YAP as a central effector. b Immunofluorescence (IF) staining showing active YAP localization in spheroid cultures of dysadherin-overexpressing (OE) or knockdown (KD) HCC cells (PLC/PRF/5 and SK-Hep1). Scale bar = 100 μm. c Heatmaps showing the expression of YAP target genes in tumors derived from the LDA experiment. d Immunoblot analysis of active and total YAP, phospho-YAP (S127), and CTGF expression in tumors derived from the LDA experiment. e Immunoblot analysis showing YAP activation status in dysadherin OE or KD cells treated with YAP knockdown (shYAP), verteporfin (VP), or constitutively active YAP (YAP5SA). f , i , l Sphere formation assays showing size and growth of spheroids under the indicated conditions (shYAP, verteporfin, or YAP5SA). Scale bar = 100 μm. g , j , m Clonogenic survival assays measuring colony-forming ability in PLC/PRF/5 or SK-Hep1 cells. h , k , n Heatmaps showing expression of CSC-associated genes in dysadherin OE or KD cells. Data are presented as means ± SEM. Statistical significance was determined by unpaired two-tailed Student’s t-tests and one-way ANOVA with Dunnett’s multiple comparison tests. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The human HCC cell lines, HepG2, SNU368, and SK-Hep1 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Republic of Korea).

Techniques: Activation Assay, Immunofluorescence, Staining, Knockdown, Expressing, Derivative Assay, Western Blot, Two Tailed Test, Comparison

Journal: Signal Transduction and Targeted Therapy

Article Title: Dysadherin/YAP axis fuels stem plasticity and immune escape in liver cancer

doi: 10.1038/s41392-025-02520-4

Figure Lengend Snippet: The dysadherin–FAK–YAP axis drives TEAD2-dependent transcription of pluripotency genes in HCC. a GSEA of DEGs from dysadherin high versus dysadherin low HCC tumors ( GSE9843 ). b , c Immunoblot analyses showing levels of phosphorylated FAK (p-FAK), total and active YAP, phospho-YAP (S127), and pluripotency transcription factors (OCT4, KLF4, MYC, SOX2) in PLC/PRF/5 and SK-Hep1 cells. Cells were subjected to dysadherin OE or KD and treated with shYAP, constitutively active YAP mutant (YAP5SA), or the FAK inhibitor PND-1186. d Immunoblot analysis showing levels of dysadherin (DYS), phosphorylated FAK (P-FAK), total FAK (T-FAK), phosphorylated LATS1/2 (P-LATS1/2), total LATS1, phosphorylated MST1/2 (P-MST1/2), and total MST1 in PLC/PRF/5 cells. Cells were subjected to dysadherin overexpression with or without a FAK inhibitor. e RT-qPCR analysis of pluripotency genes in SK-Hep1 cells expressing YAP5SA or treated with verteporfin. f Luciferase reporter assay assessing OCT4 promoter activity in dysadherin OE PLC/PRF/5 cells treated with or without verteporfin. g Venn diagram illustrating overlap of TEAD2 and TEAD4 as candidate transcriptional regulators of OCT4 from the EPD and GTRD databases. h RT-qPCR analysis of OCT4 mRNA levels in YAP5SA-expressing SK-Hep1 cells following siRNA-mediated knockdown of TEAD2 or TEAD4. i ChIP-qPCR confirming direct binding of TEAD2 to the OCT4 promoter region using seven primer sets spanning from −1400 to +9 bp relative to the transcription start site. For ChIP-Re-ChIP analysis in SK-Hep1 cells, chromatin was first immunoprecipitated with an anti-TEAD2 antibody, and the resulting material was re-immunoprecipitated with an anti-YAP antibody. PCR was performed with primers spanning the OCT4 promoter. IgG was used as a negative control. Data are presented as means ± SEM. Statistical significance was determined by unpaired two-tailed Student’s t-tests or one-way ANOVA with Dunnett’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The human HCC cell lines, HepG2, SNU368, and SK-Hep1 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Republic of Korea).

Techniques: Western Blot, Mutagenesis, Over Expression, Quantitative RT-PCR, Expressing, Luciferase, Reporter Assay, Activity Assay, Knockdown, ChIP-qPCR, Binding Assay, Immunoprecipitation, Negative Control, Two Tailed Test, Comparison

Journal: Signal Transduction and Targeted Therapy

Article Title: Dysadherin/YAP axis fuels stem plasticity and immune escape in liver cancer

doi: 10.1038/s41392-025-02520-4

Figure Lengend Snippet: The dysadherin–YAP axis modulates drug resistance and PD-L1–mediated immune evasion in HCC. a GSEA of DEGs between dysadherin high and dysadherin low tumors ( GSE9843 ), revealing enrichment of immunotherapy resistance and drug resistance signatures. b Heatmaps showing increased expression of gene sets associated with resistance to tyrosine-kinase inhibitors (left), cytostatic drugs (middle), and doxorubicin (right) in dysadherin high HCC tumors. c Cell viability and IC 50 curves of PLC/PRF/5 (dysadherin OE) and SK-Hep1 (dysadherin KD) cells treated with verteporfin or YAP5SA, in the presence of sorafenib. d Apoptosis analysis using Annexin V/PI staining in PLC/PRF/5 and SK-Hep1 cells under the same conditions as in ( c ). e Heatmap showing elevated expression of immunosuppressive genes and immune checkpoint–related transcripts in dysadherin high tumors from GSE9843 . f Immunoblot analysis of active YAP, phospho-YAP (S127), total YAP, and PD-L1 in dysadherin-modified cells treated with shYAP, verteporfin, or YAP5SA. g ChIP-qPCR showing TEAD2 binding to the PD-L1 promoter region in dysadherin-OE PLC/PRF/5 cells. ChIP-Re-ChIP analysis showing co-occupancy of TEAD2 and YAP on the PD-L1 promoter in SK-Hep1 cells. Sequential immunoprecipitation was performed first with an anti-TEAD2 antibody, followed by an anti-YAP antibody. IgG served as a negative control. h Immunofluorescence analysis of PD-L1 and PD-1 binding in dysadherin-OE and KD cells using PD-1-Fc fusion protein staining. Scale bar = 100 μm. Flow cytometry analysis of CD69 expression ( i ) and the measurement of IFN-γ secretion ( j ) in Jurkat T cells co-cultured with dysadherin-modified HCC cells, treated with verteporfin or YAP5SA under CD3/CD28 stimulation. Data are presented as means ± SEM. Statistical significance was determined by unpaired two-tailed Student’s t-tests and one-way ANOVA with Dunnett’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The human HCC cell lines, HepG2, SNU368, and SK-Hep1 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Republic of Korea).

Techniques: Expressing, Staining, Western Blot, Modification, ChIP-qPCR, Binding Assay, Immunoprecipitation, Negative Control, Immunofluorescence, Flow Cytometry, Cell Culture, Two Tailed Test, Comparison

Journal: Signal Transduction and Targeted Therapy

Article Title: Dysadherin/YAP axis fuels stem plasticity and immune escape in liver cancer

doi: 10.1038/s41392-025-02520-4

Figure Lengend Snippet: The dysadherin–YAP axis drives HCC progression and facilitates immune evasion in vivo. a Schematic overview of the humanized mouse model (Hu-NSG-SGM3) used for in vivo tumor studies. b Tumor growth of SK-Hep1 cells with or without dysadherin knockdown (DYS KD ) in the humanized mouse model. Immunofluorescence ( c , e ) and immunoblot ( d ) analysis of dysadherin, YAP, CTGF, and PD-L1 expression in SK-Hep1 tumors. Scale bar = 100 μm. f Left: Flow cytometric quantification of tumor-infiltrating CD8⁺ T cells and PD1⁺TIM3⁺ exhausted T cells. Right: Heatmap of cytotoxic and exhaustion marker gene expression profiles of T cells in DYS KD versus control tumors. g Schematic overview of the in vivo peptide treatment protocol in NSG mice. Representative tumor images ( h ) and quantification of tumor growth and weight ( i ) in SK-Hep1 xenografts treated with the dysadherin-inhibitory peptide versus vehicle (DMSO) for 28 days (n = 6 mice/group). j Immunoblotting of dysadherin, YAP activation markers, and CTGF expression in tumor tissues after peptide treatment. k Experimental design for splenic injection model of liver metastasis using PLC/PRF/5 spheroids with or without dysadherin OE and shYAP. Scale bar = 1 mm. Quantification of tumor incidence ( l ), tumor size ( m ) in the liver metastasis model. n Schematic diagram of integrin-FAK-YAP-TEAD axis-mediated stemness acquisition and immune escape in liver cancer. Created with BioRender.com. Data are presented as means ± SEM. Statistical significance was determined by unpaired two-tailed Student’s t-tests and one-way ANOVA with Dunnett’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The human HCC cell lines, HepG2, SNU368, and SK-Hep1 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Republic of Korea).

Techniques: In Vivo, Knockdown, Immunofluorescence, Western Blot, Expressing, Marker, Gene Expression, Control, Activation Assay, Injection, Two Tailed Test, Comparison