Journal: Oncology Research

Article Title: MINDY1 Induces PD-L1 Deubiquitination to Promote Immune Escape in Hepatocellular Carcinoma by the Wnt/β-Catenin Pathway

doi: 10.32604/or.2025.067638

Figure Lengend Snippet: MINDY1 promotes PD-L1 deubiquitination through Wnt/β-catenin pathway. ( A–C ) MINDY1/Vector and sh-MINDY1/sh-NC were transfected, and Western blot analysis assessed that overexpression of MINDY1 elevated β-catenin and p-GSK3β(Tyr216)/GSK3β levels in HCC cells, and knockdown of MINDY1 did the opposite. ( D – F ) Western blot measured that SKL2001 (Wnt pathway activator) elevated the activating effect of overexpression of MINDY1 on the Wnt/β-catenin pathway, while pathway inhibitor LiCl decreased this phenomenon. (G) Western blot measured that SKL2001 increased PD-L1 protein expression, whereas LiCl led to PD-L1 downregulation. (H) Western blot examined that SKL2001 promoted the deubiquitination of PD-L1 in HCC cells, while LiCl suppressed its deubiquitination. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Vector/sh-NC; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. MINDY1

Article Snippet: HCC cells transfected with MINDY1 were exposed to the Wnt inhibitor LiCl (20 mM, HY-Y0649, MedChemExpress, Monmouth Junction, NJ, USA) and incubated for 24 h, and this group was denoted as the MINDY1+LiCl group [ ].

Techniques: Plasmid Preparation, Transfection, Western Blot, Over Expression, Knockdown, Expressing

Journal: Oncology Research

Article Title: MINDY1 Induces PD-L1 Deubiquitination to Promote Immune Escape in Hepatocellular Carcinoma by the Wnt/β-Catenin Pathway

doi: 10.32604/or.2025.067638

Figure Lengend Snippet: MINDY1 hinders CD8 + T cell activation through the Wnt/β-catenin pathway. ( A ) CFSE staining confirmed that co-culture with HCC cells transfected with MINDY1 resulted in diminished proliferation of CD8 + T cells, which was further diminished by SKL2001 treatment, whereas LiCl treatment did the opposite. ( B ) Flow cytometry indicated a rise in CD8 + T cell apoptosis after they were co-cultured with HCC cells overexpressing MINDY1, with an additional rise observed by SKL2001 treatment, while LiCl treatment reduced the apoptosis rate. ( C ) The cytotoxicity of CD8 + T cells was assessed through the LDH kit. ( D , E ) Flow cytometry confirmed that co-culturing with HCC cells overexpressing MINDY1 led to a decrease in Perforin + and TNF-α + T cells. After treatment with SKL2001, this decrease was further exacerbated, while LiCl increased activated CD8 + T cells. ( F , G ) ELISA was used to detect IFN-γ and IL-2 levels. ( H ) CCK-8 assay was employed to examine the survival rates of HCC cells. n = 3. * p < 0.05, *** p < 0.001 vs. Vector; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. MINDY1

Article Snippet: HCC cells transfected with MINDY1 were exposed to the Wnt inhibitor LiCl (20 mM, HY-Y0649, MedChemExpress, Monmouth Junction, NJ, USA) and incubated for 24 h, and this group was denoted as the MINDY1+LiCl group [ ].

Techniques: Activation Assay, Staining, Co-Culture Assay, Transfection, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Plasmid Preparation