title: Epigenetic silencing of SFRP1 and SFRP5 by hepatitis B virus X protein enhances hepatoma cell tumorigenicity through Wnt signaling pathway.
abstract: Secreted frizzled-related proteins (SFRPs) are antagonists of the Wnt signaling pathway whose epigenetic downregulation have been shown to be involved in hepatocarcinogenesis. However, dysregulation of SFRPs induced by hepatitis B virus (HBV) X protein (HBx) has never been studied in HBV-related hepatocellular carcinoma (HBV-HCC). In this study, we sought to determine the clinical significance and underlying mechanism of HBx-induced SFRPs dysregulation in hepatoma cells and HBV-HCC patients. Our results showed that SFRP1 and SFRP5 expression were dramatically decreased by HBx in hepatoma cells. The repressed expression in hepatoma cells was partially rescued by a DNA methylation inhibitor and synergistically increased by a combination treatment with a histone deacetyltransferases inhibitor. In addition, we identified that SFRP1 and SFRP5 promoters were hypermethylated in both HBx-expressing hepatoma cells and HBV-HCC tissues. Downregulation of SFRP1 and SFRP5 in HBV-HCC tissues was significantly correlated with overexpression of DNA methyltransferase 1 (DNMT1) and poor tumor differentiation. HBx facilitated the binding of DNMT1 and DNMT3A to SFRP1 and SFRP5 promoters, and resulted in epigenetic silencing of SFRP1 and SFRP5. Moreover, overexpression of SFRP1, SFRP5 or RNA interference mediated silencing of DNMT1 inactivated the Wnt signaling pathway and decreased the expression levels of Wnt target genes c-Myc and CyclinD1, thus impeding HCC growth in vitro and in vivo, and regressing HBx-induced epithelial-mesenchymal transition (EMT). Our findings strongly suggest that epigenetic silencing of SFRP1 and SFRP5 by HBx allows constitutive activation of Wnt signaling pathway and hence contributes to hepatocarcinogenesis.
authors.affiliation: The Second Affiliated Hospital and the Key Laboratory of Molecular Biology of Infectious Diseases designated by the Chinese Ministry of Education, Chongqing Medical University, Chongqing, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Xie
authors.forename: Linlin
authors.initials: L
authors.lastname: Chen
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Shan
authors.forename: Xiaoliang
authors.initials: X
authors.lastname: Shan
authors.forename: Jia
authors.initials: J
authors.lastname: Tang
authors.forename: Fan
authors.initials: F
authors.lastname: Zhou
authors.forename: Qingmei
authors.initials: Q
authors.lastname: Chen
authors.forename: Huiqin
authors.initials: H
authors.lastname: Quan
authors.forename: Dan
authors.initials: D
authors.lastname: Nie
authors.forename: Wenlu
authors.initials: W
authors.lastname: Zhang
authors.forename: Ai-Long
authors.initials: AL
authors.lastname: Huang
authors.forename: Ni
authors.initials: N
authors.lastname: Tang
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002465:Cell Movement; D047369:Chromatin Immunoprecipitation; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D058750:Epithelial-Mesenchymal Transition; D005136:Eye Proteins / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006509:Hepatitis B / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006515:Hepatitis B virus; D006801:Humans; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics*; D008099:Liver / Q000378:metabolism; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008565:Membrane Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D060888:Real-Time Polymerase Chain Reaction; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D054334:Viral Regulatory and Accessory Proteins; D060449:Wnt Signaling Pathway / Q000502:physiology; D000094742:Secreted Frizzled-Related Proteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D005136:Eye Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C117636:SFRP1 protein, human; D000094742:Secreted Frizzled-Related Proteins; C118511:SFRP5 protein, human; C057628:hepatitis B virus X protein
keywords: Wnt/β-catenin signaling pathway; epigenetic silencing, SFRPs; hepatitis B virus; hepatocellular carcinoma