Journal: Chinese Medical Journal

Article Title: USP25 promotes hepatocellular carcinoma progression by interacting with TRIM21 via the Wnt/β-catenin signaling pathway

doi: 10.1097/CM9.0000000000002714

Figure Lengend Snippet: USP25 is highly expressed in HCC and is correlated with poor prognosis. (A) The expression of USP25 in pan-carcinoma as determined with the TIMER database. (B) The expression of USP25 mRNA was analyzed in HCC tissues from TCGA and ICGC databases. (C) The protein expression levels of USP25 in a normal liver cell line and six HCC cell lines. (D) The expression of USP25 in different HCC cell lines in the CCLE database. (E) Kaplan–Meier survival analysis of survival in HCC patients grouped according to USP25 expression ( n = 370). (F) The promoter methylation level of USP25 in different grades of HCC according to the analysis of patient samples. Grade 1: well differentiated; Grade 2: moderately-differentiated; Grade 3: moderately and poorly differentiated; Grade 4: poorly differentiated. (G) The expression of USP25 in HCC patients based on nodal metastasis status. * P <0.05; † P <0.001; ‡ P <0.01, § P <0.0001. ACC: Adrenocortieal carcinoma; BLCA: Bladder urothelial carcinoma; BRCA:Breast invasive carcinoma; CESC: Cervical squamous cell carcinoma; CHOL: Cholangiocarcinoma; COAD: Colon adenocarcinoma; DLBC: Lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: Esophageal carcinoma; GBM: Glioblastoma multiforme; HNSC: head and neck squamous cell carcinoma; KICH: Kidney chromophobe; KIRC: Kidney renal clear cell carcinoma; KIRP: Kidney renal papillary cell carcinoma; LAML: Acute myeloid leukemia; LGG: Brain lower grade glioma; LIHC: Liver hepatocellular carcinoma; LUAD: Lung adenocarcinoma; LUSC: Lung squamous cell carcinoma; MESO: Mesothelioma; OV: Ovarian serous cystadenocarcinoma; PAAD: Pancreatic adenocarcinoma; PCPG: Pheochromocytoma and paraganglioma; PRAD: Prostate adenocarcinoma; READ: Rectum adenocarcinoma; SARC: Sarcoma; SKCM: Skin cutaneous melanoma; STAD: Stomach adenocarcinoma; TGCT: Testicular germ cell tumors; THCA: Thyroid carcinoma; THYM: Thymoma; UCEC: Uterine corpus endometrial carcinoma; UCS: Uterine carcinosarcoma; UVM: Uveal melanoma; HCC: Hepatocellular carcinoma; ICGC: International Cancer Genome Consortium; TIMER: Tumor Immune Estimation Resource; USP25: Ubiquitin-specific peptidase 25; TCGA: The Cancer Genome Atlas; CCLE: Cancer Cell Line Encyclopedia; HR: Hazard ratio; L-O2: Normal hepatocyte cell line; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; N0, 1: Node0,1.

Article Snippet: Six human HCC cell lines (HepG2, HuH7, SMMC7721, Bel7402, MHCC97H, and PLC/PRF/5) and a normal hepatocyte cell line (L-O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Methylation, Ubiquitin Proteomics

Journal: Chinese Medical Journal

Article Title: USP25 promotes hepatocellular carcinoma progression by interacting with TRIM21 via the Wnt/β-catenin signaling pathway

doi: 10.1097/CM9.0000000000002714

Figure Lengend Snippet: USP25 promotes HCC cell proliferation in vitro. (A–C) HepG2 and MHCC97H cell lines with USP25 overexpression or knockdown were successfully established, as verified by analysis of protein and mRNA levels and fluorescence microscopy (Original magnification × 100). (D–F) Colony formation and CCK-8 assays were performed to detect cell proliferation. (G–J) DNA synthesis was measured by EdU IF staining (Original magnification × 200). * P <0.05; † P <0.001; ‡ P <0.01.CCK-8: Cell counting kit-8; HCC: Hepatocellular carcinoma; IF: Immunofluorescence; USP25: Ubiquitin-specific peptidase 25; EdU: 5-ethnyl-2-deoxyuridine; DAPI: 4′,6-Diamidino-2′-phenylindole; shUSP25: short hairpin Ubiquitin-specific peptidase 25; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Six human HCC cell lines (HepG2, HuH7, SMMC7721, Bel7402, MHCC97H, and PLC/PRF/5) and a normal hepatocyte cell line (L-O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: In Vitro, Over Expression, Knockdown, Fluorescence, Microscopy, CCK-8 Assay, DNA Synthesis, Staining, Cell Counting, Immunofluorescence, Ubiquitin Proteomics

Journal: Chinese Medical Journal

Article Title: USP25 promotes hepatocellular carcinoma progression by interacting with TRIM21 via the Wnt/β-catenin signaling pathway

doi: 10.1097/CM9.0000000000002714

Figure Lengend Snippet: USP25 promotes HCC cell migration and invasion in vitro. (A,B) A Transwell invasion assay was performed to assess the invasiveness of HepG2 and MHCC97H cells (Original magnification × 100). (C,D) A transwell migration assay was performed to analyze the migratory ability of HepG2 and MHCC97H cells (Original magnification × 100). (E–H) The scratch assay results demonstrated the migratory ability of HepG2 and MHCC97H cells (Original magnification × 40). (I,J) The expression levels of a PCNA and invasion-related proteins (MMP2 and MMP9) were detected by Western blotting. * P <0.001; † P <0.01. HCC: Hepatocellular carcinoma; PCNA: Proliferation-related protein; USP25: Ubiquitin-specific peptidase 25; MMP2: Matrix metallopeptidase 2; MMP9: Matrix metallopeptidase 9; shUSP25: short hairpin Ubiquitin-specific peptidase 25; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Six human HCC cell lines (HepG2, HuH7, SMMC7721, Bel7402, MHCC97H, and PLC/PRF/5) and a normal hepatocyte cell line (L-O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Migration, In Vitro, Transwell Invasion Assay, Transwell Migration Assay, Wound Healing Assay, Expressing, Western Blot, Ubiquitin Proteomics

Journal: Chinese Medical Journal

Article Title: USP25 promotes hepatocellular carcinoma progression by interacting with TRIM21 via the Wnt/β-catenin signaling pathway

doi: 10.1097/CM9.0000000000002714

Figure Lengend Snippet: USP25 promotes HCC growth by activating the Wnt/β-catenin signaling pathway. (A) The expression of β-catenin was observed by IF staining (Original magnification × 200). (B) The mRNA expression of components in the Wnt/β-catenin signaling pathway was measured by qRT-PCR. (C) The expression of β-catenin was observed by Western blotting of nuclear-cytoplasmic fractionation. (D, E) Western blotting was used to detect the protein expression levels of Wnt/β-catenin pathway components and downstream factors. (F, G) Western blotting analysis after XAV939 treatment of HepG2 and MHCC97H cells. * P <0.001; † P <0.01; ‡ P <0.05, shUSP25 vs . Control; § P <0.05, shUSP25+XAV939 vs . Control. HCC: Hepatocellular carcinoma; IF: Immunofluorescence; qRT-PCR: Quantitative real-time polymerase chain reactio; USP25: Ubiquitin-specific peptidase 25.DAPI: 4′,6-Diamidino-2′-phenylindole; shUSP25: Short hairpin Ubiquitin-specific peptidase 25; PCNA: Proliferating cell nuclear antigen GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; GSK3β: Glycogen synthase kinase 3 beta.

Article Snippet: Six human HCC cell lines (HepG2, HuH7, SMMC7721, Bel7402, MHCC97H, and PLC/PRF/5) and a normal hepatocyte cell line (L-O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Staining, Quantitative RT-PCR, Western Blot, Fractionation, Control, Immunofluorescence, Ubiquitin Proteomics