Analysis of the cellular uptake and nuclear delivery of insulin-like growth factor binding protein-3 in human osteosarcoma cells

Int J Cancer. 2012 Apr 1;130(7):1544-57. doi: 10.1002/ijc.26149. Epub 2011 Aug 2.

Abstract

Insulin-like growth factor (IGF) binding protein-3 (IGFBP-3) regulates cell proliferation and survival by extracellular interaction and inactivation of the growth factor IGF-I. Beyond that, IGF-independent actions mediated by intracellular IGFBP-3 including nuclear-IGFBP-3, have also been described. We here show, using both confocal and electron microscopy and cell fractionation, that the extracellular addition of IGFBP-3 to living cells results in rapid uptake and nuclear delivery of IGFBP-3, by yet partly unknown mechanisms. IGFBP-3 is internalized through a dynamin-dependent pathway, traffics through endocytic compartments and is finally delivered into the nucleus. We observed docking of IGFBP-3 containing structures to the nuclear envelope and found IGFBP-3 containing dot-like structures to permeate the nuclear envelope. In summary, our findings establish the pathway by which this tumor suppressor protein is delivered from extracellular space to the nucleus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bone Neoplasms / metabolism*
  • Cell Fractionation / methods
  • Cell Nucleus / metabolism*
  • Cytoplasm / metabolism
  • Endocytosis / physiology
  • Humans
  • Insulin-Like Growth Factor Binding Protein 3 / metabolism*
  • Insulin-Like Growth Factor Binding Protein 3 / pharmacokinetics
  • Microscopy, Confocal / methods
  • Microscopy, Electron / methods
  • Osteosarcoma / metabolism*
  • Protein Binding
  • Protein Transport / physiology*
  • Recombinant Proteins / pharmacokinetics
  • Tumor Cells, Cultured

Substances

  • IGFBP3 protein, human
  • Insulin-Like Growth Factor Binding Protein 3
  • Recombinant Proteins