miR-370 regulates ISG15 expression and influences IFN-α sensitivity in hepatocellular carcinoma cells

Cancer Biomark. 2018;22(3):453-466. doi: 10.3233/CBM-171075.

Abstract

Background: Interferon-α (IFN-α) is an adjuvant to chemotherapy and radiotherapy for hepatocellular carcinoma (HCC), but some HCC patients do not respond to treatment with IFN-α.

Methods: We performed loss-of-function and gain-of-function experiments to examine the role of ISG15 in the IFN-α sensitivity of LH86, HLCZ01, SMMC7721, and Huh7 cell lines and tumor samples.

Results: The overexpression of ISG15 reduced apoptosis in Huh7 and LH86 cells in the presence of IFN-α, whereas the shRNA-mediated knock down of ISG15 expression increased apoptosis in both Huh7 and LH86 cells. We identified a putative miR-370 target site in the 3'-UTR in the ISG15 mRNA, and the level of miR-370 expression in HCC cell lines reflected the level of IFN-α-induced apoptosis exhibited by each. Both HCC cell lines and tumor samples had significantly lower levels of miR-370 than the control cells and tissues (P< 0.05). The overexpression of miR-370 in IFN-α-treated LH86 and Huh7 cells increased apoptosis and reduced the volume of LH86- and Huh7-derived xenograft tumors in mice treated with IFN-α compared with the control tumors.

Conclusions: Our findings suggest that miR-370 functions as an HCC tumor suppressor and regulator of IFN-α sensitivity and that miR-370 might be a useful prognostic marker for HCC patients.

Keywords: Hepatocellular carcinoma cells; ISG15; interferon-α; miR-370; prognostic marker.

MeSH terms

  • 3' Untranslated Regions
  • Animals
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Carcinoma, Hepatocellular / genetics*
  • Carcinoma, Hepatocellular / immunology
  • Carcinoma, Hepatocellular / mortality
  • Carcinoma, Hepatocellular / pathology
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Survival / genetics
  • Cytokines / genetics*
  • Disease Models, Animal
  • Gene Expression Regulation, Neoplastic / drug effects*
  • Humans
  • Immunity
  • Interferon-alpha / pharmacology*
  • Liver Neoplasms / genetics*
  • Liver Neoplasms / immunology
  • Liver Neoplasms / mortality
  • Liver Neoplasms / pathology
  • Mice
  • MicroRNAs / genetics*
  • Models, Molecular
  • RNA Interference*
  • Ubiquitins / genetics*

Substances

  • 3' Untranslated Regions
  • Cytokines
  • Interferon-alpha
  • MIRN370 microRNA, human
  • MicroRNAs
  • Ubiquitins
  • ISG15 protein, human