Fluoxetine induces autophagic cell death via eEF2K-AMPK-mTOR-ULK complex axis in triple negative breast cancer

Cell Prolif. 2018 Apr;51(2):e12402. doi: 10.1111/cpr.12402. Epub 2017 Nov 1.

Abstract

Objectives: Triple negative breast cancer (TNBC) is a complex and intrinsically aggressive tumour with poor prognosis, and the discovery of targeted small-molecule drugs for TNBC treatment still remains in its infancy. In this study, we aimed to discover a small-molecule agent for TNBC treatment and illuminate its potential mechanisms.

Materials and methods: Cell viability was detected by using methylthiazoltetrazolium (MTT) assay. Electron microscopy, GFP-LC3 transfection, monodansylcadaverine staining and apoptosis assay were performed to determine Fluoxetine-induced autophagy and apoptosis. Western blotting and siRNA transfection were carried out to investigate the mechanisms of Fluoxetine-induced autophagy. iTRAQ-based proteomics analysis was used to explore the underlying mechanisms.

Results: We have demonstrated that Fluoxetine had remarkable anti-proliferative activities and induced autophagic cell death in MDA-MB-231 and MDA-MB-436 cells. The mechanism for Fluoxetine-induced autophagic cell death was associated with inhibition of eEF2K and activation of AMPK-mTOR-ULK complex axis. Further iTRAQ-based proteomics and network analyses revealed that Fluoxetine-induced mechanism was involved in BIRC6, BNIP1, SNAP29 and Bif-1.

Conclusions: These results demonstrate that Fluoxetine induces apoptosis and autophagic cell death in TNBC, which will hold a promise for the future TNBC therapy.

MeSH terms

  • AMP-Activated Protein Kinases / metabolism*
  • Apoptosis / drug effects
  • Autophagy / drug effects*
  • Breast Neoplasms / drug therapy*
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / pathology
  • Elongation Factor 2 Kinase / metabolism*
  • Female
  • Fluoxetine / pharmacology*
  • Humans
  • MCF-7 Cells
  • Multienzyme Complexes / metabolism*
  • Neoplasm Proteins / metabolism*
  • TOR Serine-Threonine Kinases / metabolism*

Substances

  • Multienzyme Complexes
  • Neoplasm Proteins
  • Fluoxetine
  • MTOR protein, human
  • EEF2K protein, human
  • TOR Serine-Threonine Kinases
  • Elongation Factor 2 Kinase
  • AMP-Activated Protein Kinases