Quantification of thrombospondin-1 secretion and expression of alphavbeta3 and alpha3beta1 integrins and syndecan-1 as cell-surface receptors for thrombospondin-1 in malignant glioma cells

J Neurooncol. 2004 Dec;70(3):309-17. doi: 10.1007/s11060-004-9167-1.

Abstract

Malignant glioma cells secrete thrombospondin-1 (TSP-1) which participates in the motility of glioma cells, and binds to cell surface alphavbeta3 and alpha3beta1 integrins, and syndecan-1. This study evaluated the amount of TSP-1 secretion from malignant glioma cells, and the expression of alphavbeta3 and alpha3beta1 integrins, and syndecan-1. The amounts of TSP-1 in the supernatants from 10 malignant glioma cell lines and eight non-glioma malignant tumor cell lines were measured by enzyme-linked immunosorbent assay. Expression of alphavbeta3 and alpha3beta1 integrins, and syndecan-1 were examined by flow cytometry. The amounts of TSP-1 secreted by malignant glioma cells were 43 to 2431 ng/l x 10(6) cells/24 h (mean +/- SD = 626 +/- 792). Seven of 10 glioma cell lines secreted more than 100 ng of TSP-1 and three of these cell lines secreted more than 1 microg. Seven of eight non-glioma cell lines secreted less than 100 ng of TSP-1. All glioma cell lines expressed alpha3beta1 integrin and syndecan-1, and seven of 10 glioma cell lines expressed alphavbeta3 integrin. Treatment of the glioma cell lines with TGF-beta2 did not change the expression of alphavbeta3 integrin. These results suggest that malignant glioma cells secrete high levels of TSP-1, which may be important in the migration of glioma cells via interactions with alphavbeta3 and alpha3beta1 integrins, and syndecan-1.

MeSH terms

  • Brain Neoplasms / metabolism*
  • Cell Line, Tumor
  • Cell Movement / physiology
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Glioma / metabolism*
  • Humans
  • Integrin alpha3beta1 / biosynthesis*
  • Integrin alphaVbeta3 / biosynthesis*
  • Membrane Glycoproteins / biosynthesis*
  • Proteoglycans / biosynthesis*
  • Syndecan-1
  • Syndecans
  • Thrombospondin 1 / metabolism*

Substances

  • Integrin alpha3beta1
  • Integrin alphaVbeta3
  • Membrane Glycoproteins
  • Proteoglycans
  • SDC1 protein, human
  • Syndecan-1
  • Syndecans
  • Thrombospondin 1