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Cell surface heparan sulfate proteoglycans are involved in the extracellular Hsp90-stimulated migration and invasion of cancer cells

  • Anastasiya V. Snigireva
  • Veronika V. Vrublevskaya
  • Yuri Y. Skarga
  • Oleg S. MorenkovEmail author
Original Paper

Abstract

The extracellular heat shock protein 90 (Hsp90) is known to participate in cell migration and invasion. Recently, we have shown that cell surface heparan sulfate proteoglycans (HSPGs) are involved in the binding and anchoring of extracellular Hsp90 to the plasma membrane, but the biological relevance of this finding was unclear. Here, we demonstrated that the digestion of heparan sulfate (HS) moieties of HSPGs with a heparinase I/III blend and the metabolic inhibition of the sulfation of HS chains by sodium chlorate considerably impair the migration and invasion of human glioblastoma A-172 and fibrosarcoma HT1080 cells stimulated by extracellular native Hsp90. Heparin, a polysaccharide closely related to HS, also reduced the Hsp90-stimulated migration and invasion of cells. Phorbol 12-myristate 13-acetate, an intracellular inducer of cell motility bypassing the ligand activation of receptors, restored the basal migration of heparinase- and chlorate-treated cells almost to the control level, suggesting that the cell motility machinery was insignificantly affected in cells with degraded and undersulfated HS chains. On the other hand, the downstream phosphorylation of AKT in response to extracellular Hsp90 was substantially impaired in heparinase- and chlorate-treated cells as compared to untreated cells. Taken together, our results demonstrated for the first time that cell surface HSPGs play an important role in the migration and invasion of cancer cells stimulated by extracellular Hsp90 and that plasma membrane-associated HSPGs are required for the efficient transmission of signal from extracellular Hsp90 into the cell.

Keywords

Cell surface HSPGs Extracellular Hsp90 Hsp90-stimulated cell migration Hsp90-stimulated cell invasion 

Notes

Acknowledgments

We thank A.O. Shepelyakovskaya for the help with flow cytometry.

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflict of interest.

Supplementary material

12192_2018_955_MOESM1_ESM.pdf (95 kb)
Fig. S1 Purified native mouse Hsp90 was subjected to SDS-PAGE (a) and Western blot analysis with Hsp90α- and Hsp90β-specific antibodies (b). (PDF 95 kb)
12192_2018_955_MOESM2_ESM.pdf (44 kb)
Fig. S2 The stimulation of migration and invasion of cells by native Hsp90s from different animal species (a) and the analysis of cytotoxicity and antiproliferative activity of native mouse Hsp90 (b). (a) The migration and invasion of cells were evaluated in the transwell migration/invasion assays in the absence and presence of mouse, swine, and bovine Hsp90 (50 μg/ml). The migration and invasion of cells were expressed in percent relative to that of control cells without Hsp90. Each bar represents the mean ± SD (n = 5). *p < 0.05 and **p < 0.01 indicate statistically significant differences. The representative results are shown. (b) For the determination of cytotoxicity, A-172 and HT1080 cells were grown in wells of 96-well plates till confluence, then mouse Hsp90 diluted at different concentrations in DMEM-FBS was added to cells, and the plates were incubated for 48 h at 37 °C in an atmosphere of 5% CO2. After incubation, 50 μl of an MTT solution (5 mg/ml) was added to each well, and the plates were incubated at 37 °C for 2 h. The medium was removed from the wells, precipitated crystals were dissolved with DMSO, and the optical density was measured at 495 nm (OD495). To determine the antiproliferative activity, the cells were placed in wells of 96-well plates (5.0–7.0 × 103 cells per well) in DMEM-FBS containing native mouse Hsp90 at different concentrations and incubated for 48 h. Staining with MTT and measurements of OD495 values were performed as described above. The absorbance of wells expressed in percent is presented, and OD495 values of control wells without Hsp90 is taken as 100%. Each figure represents the mean ± SD (n = 5). (PDF 43 kb)

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Copyright information

© Cell Stress Society International 2019

Authors and Affiliations

  • Anastasiya V. Snigireva
    • 1
  • Veronika V. Vrublevskaya
    • 1
  • Yuri Y. Skarga
    • 1
  • Oleg S. Morenkov
    • 1
    Email author
  1. 1.Laboratory of Cell Culture and Cell Engineering, Institute of Cell BiophysicsRussian Academy of SciencesPushchinoRussia

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