Characterization of Heparan Sulfate Proteoglycan-positive Recycling Endosomes Isolated from Glioma Cells

Cancer Genomics Proteomics. 2016 11-12;13(6):443-452. doi: 10.21873/cgp.20007.

Abstract

Background: Heparan sulfate proteoglycans (HSPGs)-dependent endocytic events have been involved in glioma progression. Thus, comprehensive understanding of the intracellular trafficking complexes formed in presence of HSPGs would be important for development of glioma treatments.

Materials and methods: Subcellular fractionation was used to separate vesicles containing HSPGs from the rat C6 glioma cell line. Isolated HSPG-positive vesicles were further characterized with liquid chromatography-mass spectrometry.

Results: The HSPG-positive vesicular fractions, distinct from plasma membrane-derived material, were enriched in endocytic marker, Rab11. Proteomic analysis identified more than two hundred proteins to be associated with vesicular membrane, among them, over eighty were related to endosomal uptake, recycling or vesicular transport.

Conclusion: Part of HSPGs in glioma cells is internalized through clathrin-dependent endocytosis and undergo recycling. The development of compounds regulating HSPG-mediated trafficking will likely enable design of effective glioma treatment.

Keywords: Proteomics; Rab11; heparan sulfate proteoglycan; transport vesicle.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Clathrin / genetics
  • Endocytosis / genetics
  • Endosomes / metabolism
  • Endosomes / pathology
  • Glioma / genetics
  • Glioma / metabolism*
  • Glioma / pathology
  • Heparan Sulfate Proteoglycans / genetics
  • Heparan Sulfate Proteoglycans / metabolism*
  • Humans
  • Proteomics
  • Rats
  • Transport Vesicles / metabolism*
  • Transport Vesicles / pathology
  • rab GTP-Binding Proteins / biosynthesis*
  • rab GTP-Binding Proteins / genetics

Substances

  • Clathrin
  • Heparan Sulfate Proteoglycans
  • rab11 protein
  • rab GTP-Binding Proteins