PEITC-mediated inhibition of mRNA translation is associated with both inhibition of mTORC1 and increased eIF2α phosphorylation in established cell lines and primary human leukemia cells

Oncotarget. 2016 Nov 15;7(46):74807-74819. doi: 10.18632/oncotarget.11655.

Abstract

Increased mRNA translation drives carcinogenesis and is an attractive target for the development of new anti-cancer drugs. In this work, we investigated effects of phenethylisothiocyanate (PEITC), a phytochemical with chemopreventive and anti-cancer activity, on mRNA translation. PEITC rapidly inhibited global mRNA translation in human breast cancer-derived MCF7 cells and mouse embryonic fibroblasts (MEFs). In addition to the known inhibitory effects of PEITC on mTORC1 activity, we demonstrate that PEITC increased eIF2α phosphorylation. PEITC also increased formation of stress granules which are typically associated with eIF2α phosphorylation and accumulation of translationally stalled mRNAs. Analysis of genetically modified MEFs demonstrated that optimal inhibition of global mRNA translation by PEITC was dependent on eIF2α phosphorylation, but not mTORC1 inhibition. We extended this study into primary leukemic B cells derived from patients with chronic lymphocytic leukaemia (CLL). CLL cells were stimulated in vitro with anti-IgM to mimic binding of antigen, a major driver of this leukemia. In CLL cells, PEITC increased eIF2α phosphorylation, inhibited anti-IgM-induced mTORC1 activation and decreased both basal and anti-IgM-induced global mRNA translation. PEITC also inhibited transcription and translation of MYC mRNA and accumulation of the MYC oncoprotein, in anti-IgM-stimulated cells. Moreover, treatment of CLL cells with PEITC and the BTK kinase inhibitor ibrutinib decreased anti-IgM-induced translation and induced cell death to a greater extent than either agent alone. Therefore, PEITC can inhibit both global and mRNA specific translation (including MYC) via effects on multiple regulatory pathways. Inhibition of mRNA translation may contribute to the chemopreventive and anti-cancer effects of PEITC.

Keywords: MYC; eIF2α; mRNA translation; mTORC1; phenethylisothiocyanate.

MeSH terms

  • Antibodies, Anti-Idiotypic / pharmacology
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Dose-Response Relationship, Drug
  • Eukaryotic Initiation Factor-2 / metabolism*
  • Gene Expression Regulation, Leukemic / drug effects
  • Genes, myc
  • Humans
  • Isothiocyanates / pharmacology*
  • Leukemia / genetics*
  • Leukemia / metabolism*
  • Leukemia, Lymphocytic, Chronic, B-Cell / genetics
  • Leukemia, Lymphocytic, Chronic, B-Cell / metabolism
  • MCF-7 Cells
  • Mechanistic Target of Rapamycin Complex 1 / metabolism*
  • Phosphorylation / drug effects
  • Protein Biosynthesis / drug effects*
  • RNA, Messenger / genetics
  • Receptors, Antigen, B-Cell / metabolism
  • Stress, Physiological
  • Transcription, Genetic / drug effects

Substances

  • Antibodies, Anti-Idiotypic
  • Eukaryotic Initiation Factor-2
  • Isothiocyanates
  • RNA, Messenger
  • Receptors, Antigen, B-Cell
  • anti-IgM
  • phenethyl isothiocyanate
  • Mechanistic Target of Rapamycin Complex 1