Enhanced nuclear factor-kappa B-associated Wnt-1 expression in hepatitis B- and C-related hepatocarcinogenesis: identification by functional proteomics

Abstract

SummaryChronic infections with hepatitis B and C viruses (HBV and HCV) are etiologically linked to hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). Both viruses may induce activation of nuclear factor-kappa B (NF-jB) in hepatocytes that plays a crucial role in the regulation of cell growth and apoptosis. Functional proteomics analysis of proteins associated with NF-jB signaling complexes in both viruses-related HCC tumor and non-tumor tissues may disclose possible common mechanisms in hepatocarcinogenesis. By functional proteomics, we analyzed proteins associated with NF-jB-signaling complexes in four-paired human HCC tumor and non-tumor tissues from HBV-and HCV-infected patients, respectively, and in onepaired tissue with dual viral infection. The quantity of NF-jB-associated proteins was semi-quantitatively measured by protein spot intensity on the gels of two-dimensional polyacrylamide gel electrophoresis. The results showed that overexpression of NF-jB-associated Wnt-1 protein in tumor part was detected in the majority of HBV-and HCV-infected HCC samples. These data suggest that enhanced expression of NF-jB-associated Wnt-1 protein might be a mechanism of hepatocarcinogenesis common to HBV-and HCV-infected patients. NF-jB signaling pathway and Wnt-1 protein could be potential targets for designing highly effective therapeutic agents in treating HCC and for chemoprevention of hepatocarcinogenesis.Abbreviations: m/z -mass-to-charge ratio; 2-DE -two-dimensional polyacrylamide gel electrophoresis; EMSA -electrophoretic mobility shift assay; HBV -hepatitis B virus; HCC -hepatocellular carcinoma; HCV -hepatitis C virus; HSP -heat shock protein; MALDI-Q-TOF -matrix-assisted laser desorption/ ionization-quadrupole-time-of-flight; MS -mass spectrometry; NF-jB -nuclear factor-kappa B

Anti-sense morpholino oligonucleotide assay shows critical involvement for NF-κB activation in the production of Wnt-1 protein by HepG2 cells: oncology implications

“…It shows that the Wnt-1-mediated survival of PC12 cells is dependent on NF-jB activation, and that stable expression of Wnt-1 increases NF-jB activity [19]. This link can be indirectly supported in our recent report that functional proteomics identifies enhanced expression of NF-jBassociated Wnt-1 protein in human hepatocellular carcinoma (HCC) tissues [20]. However, a direct evidence of NF-jB activation relating to the production of Wnt-1 protein in human cells remains elusive.…”

“…Inc., South San Francisco, CA , USA) was performed by the procedures described previously [20]. The quantity of Wnt-1 protein production was semi-quantitatively estimated on the immunoblot films with ImageMaster TotalLab, Version 2.01 (Amersham Parmacia Biotech, NJ, USA), and data were estimated and expressed as relative density to HepG2 cell control.…”

Wnt-1 Protein as a Prognostic Biomarker for Hepatitis B–Related and Hepatitis C–Related Hepatocellular Carcinoma after Surgery

“…Western blotting of tumor and nontumor proteins with biotin-conjugated rabbit anti-human WNT1 polyclonal antibody (Zymed Laboratories, Inc.) was conducted by the procedures described previously (18). The amount of Wnt-1 protein was semiquantitatively measured on the immunoblot films with ImageMaster TotalLab version 2.01 (Amersham Parmacia Biotech), and all data were expressed as tumor/ nontumor ratio.…”

“…[12][13][14][15][16]. Functional proteomics and cDNA microarray studies have shown up-regulation of Wnt-1 in HBV-and HCVrelated HCC tissues and cell lines (17)(18)(19). Wnt-1 protein processing and secretion have been extensively characterized for the 36-kDa unglycosylated form as well as sequentially glycosylated forms of 38-, 40-, and 42-kDa forms.…”

“…In a mouse model of breast cancer, Wnt-1 is shown to be fundamental for cancer metastasis progression (23). Enhanced expression of nuclear factor-nBassociated Wnt-1 protein has been identified in a large proportion of HCC specimens with HBV and/or HCV infection (18). The nuclear factor-nB -P50 subunit has been shown to bind to the promoter of the Wnt-1 gene by chromatin immunoprecipitation assay (24).…”