LMP1 Increases Expression of NADPH Oxidase (NOX) and Its Regulatory Subunit p22 in NP69 Nasopharyngeal Cells and Makes Them Sensitive to a Treatment by a NOX Inhibitor

PLoS One. 2015 Aug 5;10(8):e0134896. doi: 10.1371/journal.pone.0134896. eCollection 2015.

Abstract

Oxidative stress is thought to contribute to cancer development. Epstein-Barr virus (EBV) and its encoded oncoprotein, latent membrane protein 1 (LMP1), are closely associated with the transformation of nasopharyngeal carcinoma (NPC) and Burkitt's lymphoma (BL). In this study, we used LMP1-transformed NP cells and EBV-related malignant cell lines to assess the effects of LMP1 on reactive oxygen species (ROS) accumulation and glycolytic activity. Using NPC tissue samples and a tissue array to address clinical implications, we report that LMP1 activates NAD(P)H oxidases to generate excessive amount of ROS in EBV-related malignant diseases. By evaluating NAD(P)H oxidase (NOX) subunit expression, we found that the expression of the NAD(P)H oxidase regulatory subunit p22phox was significantly upregulated upon LMP1-induced transformation. Furthermore, this upregulation was mediated by the c-Jun N-terminal kinase (JNK) pathway. In addition, LMP1 markedly stimulated anaerobic glycolytic activity through the PI3K/Akt pathway. Additionally, in both NPC cells and tissue samples, p22phox expression correlated with LMP1 expression. The NAD(P)H oxidase inhibitor diphenyleneiodonium (DPI) also exerted a marked cytotoxic effect in LMP1-transformed and malignant cells, providing a novel strategy for anticancer therapy.

MeSH terms

  • Carcinoma
  • Cell Line
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cell Survival / genetics
  • Cell Transformation, Neoplastic / genetics*
  • Cell Transformation, Neoplastic / metabolism
  • Enzyme Inhibitors / pharmacology
  • Epithelial Cells / metabolism
  • Epithelial Cells / pathology
  • Epithelial Cells / virology
  • Gene Expression Regulation, Neoplastic
  • Glycolysis
  • Herpesvirus 4, Human / genetics*
  • Herpesvirus 4, Human / metabolism
  • Herpesvirus 4, Human / physiology
  • Host-Pathogen Interactions / genetics
  • Humans
  • Immunoblotting
  • Immunohistochemistry
  • JNK Mitogen-Activated Protein Kinases / metabolism
  • NADPH Oxidases / antagonists & inhibitors
  • NADPH Oxidases / genetics*
  • NADPH Oxidases / metabolism
  • Nasopharyngeal Carcinoma
  • Nasopharyngeal Neoplasms / genetics
  • Nasopharyngeal Neoplasms / metabolism
  • Nasopharyngeal Neoplasms / virology
  • Nasopharynx / metabolism
  • Nasopharynx / pathology
  • Nasopharynx / virology
  • Onium Compounds / pharmacology
  • Phosphatidylinositol 3-Kinases / metabolism
  • Proto-Oncogene Proteins c-akt / metabolism
  • Reactive Oxygen Species / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Viral Matrix Proteins / genetics*
  • Viral Matrix Proteins / metabolism

Substances

  • EBV-associated membrane antigen, Epstein-Barr virus
  • Enzyme Inhibitors
  • Onium Compounds
  • Reactive Oxygen Species
  • Viral Matrix Proteins
  • diphenyleneiodonium
  • NADPH Oxidases
  • CYBA protein, human
  • Phosphatidylinositol 3-Kinases
  • Proto-Oncogene Proteins c-akt
  • JNK Mitogen-Activated Protein Kinases

Grants and funding

The authors received no specific funding for this work.