Influence of miR-520e-mediated MAPK signalling pathway on HBV replication and regulation of hepatocellular carcinoma cells via targeting EphA2

J Viral Hepat. 2019 Apr;26(4):496-505. doi: 10.1111/jvh.13048. Epub 2019 Feb 18.

Abstract

We determined the role of miR-520e in the replication of hepatitis B virus (HBV) and the growth of hepatocellular carcinoma (HCC) cells. MiR-520e and EPH receptor A2 (EphA2) in HBV-positive HCC tissues and cells were detected, and we studied the impact of miR-520e and the EphA2 receptor in cellular and murine HBV replication models. We find that MiR-520e was upregulated and EphA2 was downregulated in HBV-positive HCC tissues and cells. MiR-520e was decreased in Huh7-X and HepG2-X cells in which HBx was stably expressed, but was dose-dependently elevated after interfering with HBx. Additionally, miR-520e mimic and si-EphA2 groups were reduced in association with increases in HBV DNA content, HBsAg and HBeAg levels, cell proliferation and were enhanced in the expressions of EphA2, p-p38MAPK/p38MAPK, phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2)/ERK1/2 and cell apoptosis. Furthermore, si-EphA2 reversed the promotion effect of miR-520e inhibitor on HBV replication and tumour cell growth. Upregulating miR-520e in rAAV8-1.3HBV-infected mouse resulted in reduced EphA2 in liver tissues and HBV DNA content in serum. We find that MiR-520e was decreased in HBV-positive HCC, while overexpression of miR-520e blocked p38MAPK and ERK1/2 signalling pathways by an inhibitory effect on EphA2 and ultimately reduced HBV replication and inhibited tumour cell growth. These data indicate a role for miR-520e in the regulation of HBV replication.

Keywords: EPH receptor A2; hepatitis B virus; hepatocellular carcinoma; miR-520e.

MeSH terms

  • Adult
  • Animals
  • Apoptosis
  • Carcinoma, Hepatocellular / metabolism
  • Carcinoma, Hepatocellular / pathology
  • Carcinoma, Hepatocellular / virology*
  • Cell Line, Tumor
  • Cell Proliferation
  • Ephrin-A2 / genetics
  • Ephrin-A2 / metabolism*
  • Female
  • Gene Expression
  • Gene Expression Regulation, Neoplastic
  • Gene Silencing
  • Hep G2 Cells
  • Hepatitis B Surface Antigens / metabolism
  • Hepatitis B e Antigens / metabolism
  • Hepatitis B virus / physiology*
  • Humans
  • Liver Neoplasms / metabolism
  • Liver Neoplasms / pathology
  • Liver Neoplasms / virology*
  • MAP Kinase Signaling System* / genetics
  • Male
  • Mice
  • MicroRNAs / antagonists & inhibitors
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Middle Aged
  • Receptor, EphA2
  • Trans-Activators / metabolism
  • Viral Regulatory and Accessory Proteins
  • Virus Replication*

Substances

  • EPHA2 protein, human
  • Ephrin-A2
  • Hepatitis B Surface Antigens
  • Hepatitis B e Antigens
  • MIRN520 microRNA, human
  • MicroRNAs
  • Trans-Activators
  • Viral Regulatory and Accessory Proteins
  • hepatitis B virus X protein
  • Receptor, EphA2