Abstract
Loss of E-cadherin is associated with acquisition of metastatic capacity. Numerous studies suggest histone deacetylation and/or hypermethylation of CpG islands in E-cadherin gene (
CDH1
) are major mechanisms responsible for E-cadherin silencing in different tumors and cancer cell lines. The Hepatitis B virus (HBV) encoded X antigen, HBx, contributes importantly to the development of hepatocellular carcinoma (HCC) using multiple mechanisms. Experiments were designed to test if in addition to
CDH1
hypermethylation HBx promotes epigenetic modulation of E-cadherin transcriptional activity through histone deacetylation and miR-373. The relationships between HBx, E-cadherin, mSin3A, Snail-1 and miR-373 were evaluated in HBx expressing (HepG2X) and control (HepG2CAT) cells by western blotting, immunoprecipitation, chromatin immunoprecipitation as well as by immunohistochemical staining of liver and tumor tissue sections from HBV infected patients. In HepG2X cells, decreased levels of E-cadherin and elevated levels of mSin3A and Snail-1 were detected. Reciprocal immunoprecipitation with anti-HBx and anti-mSin3A demonstrated mutual binding. Further, HBx-mSin3A co-localization was detected by immunofluorescent staining. HBx down-regulated E-cadherin expression by the recruitment of the mSin3A/HDAC complex to the Snail-binding sites in human
CDH1
. Histone deacetylation inhibition by Trichostatin A treatment restored E-cadherin expression. Mir-373, a positive regulator of E-cadherin expression, was down-regulated by HBx in HepG2X cells and tissue sections from HBV infected patients. Thus, histone deacetylation of
CDH1
and down-regulation of miR-373, together with the previously demonstrated hyper-methylation of
CDH1
by HBx, may be important for the understanding of HBV-related carcinogenesis.