STIM1/ORAI1-mediated Ca2+ Influx Regulates Enolase-1 Exteriorization

J Biol Chem. 2015 May 8;290(19):11983-99. doi: 10.1074/jbc.M114.598425. Epub 2015 Mar 24.


Tumor cells use broad spectrum proteolytic activity of plasmin to invade tissue and form metastatic foci. Cell surface-associated enolase-1 (ENO-1) enhances plasmin formation and thus participates in the regulation of pericellular proteolysis. Although increased levels of cell surface bound ENO-1 have been described in different types of cancer, the molecular mechanism responsible for ENO-1 exteriorization remains elusive. In the present study, increased ENO-1 protein levels were found in ductal breast carcinoma and on the cell surface of highly metastatic breast cancer cell line MDA-MB-231. Elevated cell surface-associated ENO-1 expression correlated with augmented MDA-MB-231 cell migratory and invasive properties. Exposure of MDA-MB-231 cells to LPS potentiated translocation of ENO-1 to the cell surface and its release into the extracellular space in the form of exosomes. These effects were independent of de novo protein synthesis and did not require the classical endoplasmic reticulum/Golgi pathway. LPS-triggered ENO-1 exteriorization was suppressed by pretreatment of MDA-MB-231 cells with the Ca(2+) chelator BAPTA or an inhibitor of endoplasmic reticulum Ca(2+)-ATPase pump, cyclopiazonic acid. In line with these observations, the stromal interaction molecule (STIM) 1 and the calcium release-activated calcium modulator (ORAI) 1-mediated store-operated Ca(2+) entry were found to regulate LPS-induced ENO-1 exteriorization. Pharmacological blockage or knockdown of STIM1 or ORAI1 reduced ENO-1-dependent migration of MDA-MB-231 cells. Collectively, our results demonstrate the pivotal role of store-operated Ca(2+) channel-mediated Ca(2+) influx in the regulation of ENO-1 exteriorization and thus in the modulation of cancer cell migratory and invasive properties.

Keywords: calcium; cancer; enolase-1; inflammation; membrane trafficking; stromal interaction molecule 1 (STIM1).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Tumor / metabolism*
  • Biotinylation
  • Calcium / metabolism*
  • Calcium Channels / metabolism*
  • Cell Line, Tumor
  • Cell Membrane / enzymology*
  • Cell Movement
  • Cell Proliferation
  • Chelating Agents / chemistry
  • DNA-Binding Proteins / metabolism*
  • Egtazic Acid / analogs & derivatives
  • Egtazic Acid / chemistry
  • Exosomes / metabolism
  • Gene Expression Regulation, Enzymologic
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Indoles / chemistry
  • Inflammation
  • MCF-7 Cells
  • Membrane Proteins / metabolism*
  • Neoplasm Invasiveness
  • Neoplasm Metastasis
  • Neoplasm Proteins / metabolism*
  • Neoplasms / enzymology*
  • ORAI1 Protein
  • Phosphopyruvate Hydratase / metabolism*
  • Stromal Interaction Molecule 1
  • Trichloroacetic Acid / chemistry
  • Tumor Suppressor Proteins / metabolism*


  • Biomarkers, Tumor
  • Calcium Channels
  • Chelating Agents
  • DNA-Binding Proteins
  • Indoles
  • Membrane Proteins
  • Neoplasm Proteins
  • ORAI1 Protein
  • ORAI1 protein, human
  • STIM1 protein, human
  • Stromal Interaction Molecule 1
  • Tumor Suppressor Proteins
  • Egtazic Acid
  • Trichloroacetic Acid
  • ENO1 protein, human
  • Phosphopyruvate Hydratase
  • 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid
  • Calcium
  • cyclopiazonic acid