HBx induced AFP receptor expressed to activate PI3K/AKT signal to promote expression of Src in liver cells and hepatoma cells

BMC Cancer. 2015 May 6:15:362. doi: 10.1186/s12885-015-1384-9.

Abstract

Background: Hepatitis B virus (HBV)-X protein(HBx) is a transactivator of host several cellular genes including alpha-fetoprotein(AFP) and AFP receptor(AFPR) which contributes to HBV-associated tumor development. The expression of AFP/AFPR are correlated with hepatocellular carcinoma(HCC)-initial cells. But the role of AFP and AFPR in promoting occurrence of HBV-related HCC were still unclear.

Methods: A total of 71 clinical patients' liver specimens, normal human liver cells L-02 and HCC cell lines, PLC/PRF/5 were selected for analyzing the effects of HBx on expression of AFP, AFPR and Src. The expression of goal proteins were detected by Immunohistochemical stained and Western blotting; HBx-expressed vectors were constructed and transfected into L-02 cells, laser confocal microscopy was applied to observe expression and location of AFP, AFPR and Src in the normal liver cells and HCC cells, soft agar colony formation assay was used to observe colonies formed of the cells.

Results: We confirmed HBx gives preference to promote the expression of AFP and AFPR; HBx priors to up-regulate the expression of AFPR and AFP in L-02 cells and in normal liver specimens; AFPR signal been able to stimulate Src expression. The results also indicated that phosphatidylinositol 3-kinase(PI3K) inhibitors Ly294002 and GDC0941 effectively suppress AFPR mediated up-regulation expression of Src in AFPR positive HCC lines.

Conclusions: HBx priors to drive the expression of AFP and AFPR to promote expression of Src in normal liver cells and hepatoma cells; AFP and AFPR maybe play pivotal role in HBV-related hepatocarcinogenesis; Targeting AFPR is an available therapeutic strategy of HCC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Carcinoma, Hepatocellular / genetics
  • Carcinoma, Hepatocellular / metabolism*
  • Carcinoma, Hepatocellular / virology
  • Cell Line, Tumor
  • Female
  • Gene Expression
  • Gene Expression Regulation, Neoplastic
  • Hepatitis B / genetics
  • Hepatitis B / metabolism
  • Humans
  • Liver / metabolism
  • Liver / pathology
  • Liver / virology
  • Liver Neoplasms / genetics
  • Liver Neoplasms / metabolism*
  • Liver Neoplasms / virology
  • Male
  • Middle Aged
  • Phosphatidylinositol 3-Kinases / metabolism
  • Proto-Oncogene Proteins c-akt / metabolism
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • Signal Transduction
  • Trans-Activators / physiology*
  • Transcriptional Activation
  • Viral Regulatory and Accessory Proteins
  • Young Adult
  • alpha-Fetoproteins / genetics
  • alpha-Fetoproteins / metabolism
  • src-Family Kinases / genetics
  • src-Family Kinases / metabolism

Substances

  • AFP protein, human
  • NR5A2 protein, human
  • Receptors, Cytoplasmic and Nuclear
  • Trans-Activators
  • Viral Regulatory and Accessory Proteins
  • alpha-Fetoproteins
  • hepatitis B virus X protein
  • src-Family Kinases
  • Proto-Oncogene Proteins c-akt