High load hepatitis B virus replication inhibits hepatocellular carcinoma cell metastasis through regulation of epithelial-mesenchymal transition

Abstract

Objectives: The aims of this study were to investigate the effect of hepatitis B virus (HBV) replication on the metastatic ability of hepatocellular carcinoma (HCC) cells and to explore a potential mechanism from the perspective of epithelial-mesenchymal transition (EMT).

Methods: Two short-interfering RNAs (siRNAs) against the HBV S gene were used to inhibit HBV replication in HepG2.2.15 cells. To evaluate the level of HBV replication and interference efficiency, HBV antigen and HBV DNA were detected by ELISA and quantitative PCR (Q-PCR). Invasion and metastatic abilities were compared between different groups by wound healing and trans-well assays. Immunofluorescent staining and Western blotting were utilized to detect EMT markers.

Results: Both siRNAs effectively inhibited HBV replication in HepG2.2.15 cells. Compared to control HepG2.2.15 cells, cells transfected with the siRNAs showed characteristics of the mesenchymal phenotype and augmented their ability to invade and metastasize. Inhibition of HBV replication suppressed E-cadherin and induced a switch to vimentin expression. Western blots confirmed the decrease in E-cadherin expression. The level of E-cadherin expression was also lower in HepG2 cells than in HepG2.2.15 cells.

Conclusions: siRNAs were able to effectively inhibit HBV replication in vitro. A high load of HBV replication may inhibit the invasion and metastatic ability of HCC cells by reversing the EMT process.

Keywords: EMT; HBV; HepG2.2.15; Metastasis; siRNA.