Involvement of Ext1 and heparanase in migration of mouse FBJ osteosarcoma cells

Yinan Wang, Xiaoyan Yang, Sadako Yamagata, Tatsuya Yamagata, Toshinori Sato

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

To know the involvement of glycosaminoglycans (GAGs) in the metastasis of mouse FBJ osteosarcoma cells, N α -lauroyl-O-(β-d- xylopyranosyl)-l-serinamide (Xyl-Ser-C12), which initiates elongation of GAG chains using the glycan biosynthesis system in cells, was administered to FBJ cells with different metastatic capacities. Production of glycosylated products derived from Xyl-Ser-C12, especially heparan sulfate (HS) GAG-type oligosaccharides such as GalNAc-GlcA-GlcNAc-GlcA-Gal-Gal-Xyl-Ser-C12, was indicated in poorly metastatic FBJ-S1 cells more than in highly metastatic FBJ-LL cells by LC-MS. The results of RT-PCR revealed that HS synthases, Ext1 and Ext2, were expressed in FBJ-S1 cells more than in FBJ-LL cells. Furthermore, siRNA against Ext1 suppressed the expression of HS and enhanced the motility of FBJ-S1 cells. In addition, the expression of heparanase (HPSE) was enhanced in Ext-1-knockdown FBJ-S1 cells, and responsible for the increase in cell motility caused by the down-regulation of Ext1 expression. Our data provide the first evidence that Ext1 regulates the expression of HPSE and also indicated that levels of Ext1 and HPSE influenced the motility of FBJ cells.

Original languageEnglish
Pages (from-to)63-72
Number of pages10
JournalMolecular and Cellular Biochemistry
Volume373
Issue number1-2
DOIs
Publication statusPublished - 2013 Jan

Fingerprint

Heparitin Sulfate
Osteosarcoma
Glycosaminoglycans
Biosynthesis
Oligosaccharides
Small Interfering RNA
Polysaccharides
Elongation
Cell Movement
heparanase
Down-Regulation
Neoplasm Metastasis
Polymerase Chain Reaction

Keywords

  • Cell motility
  • Ext1
  • FBJ osteosarcoma
  • Glycosaminoglycan
  • Heparan sulfate
  • Heparanase
  • Saccharide primer

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Involvement of Ext1 and heparanase in migration of mouse FBJ osteosarcoma cells. / Wang, Yinan; Yang, Xiaoyan; Yamagata, Sadako; Yamagata, Tatsuya; Sato, Toshinori.

In: Molecular and Cellular Biochemistry, Vol. 373, No. 1-2, 01.2013, p. 63-72.

Research output: Contribution to journalArticle

Wang, Yinan ; Yang, Xiaoyan ; Yamagata, Sadako ; Yamagata, Tatsuya ; Sato, Toshinori. / Involvement of Ext1 and heparanase in migration of mouse FBJ osteosarcoma cells. In: Molecular and Cellular Biochemistry. 2013 ; Vol. 373, No. 1-2. pp. 63-72.
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AB - To know the involvement of glycosaminoglycans (GAGs) in the metastasis of mouse FBJ osteosarcoma cells, N α -lauroyl-O-(β-d- xylopyranosyl)-l-serinamide (Xyl-Ser-C12), which initiates elongation of GAG chains using the glycan biosynthesis system in cells, was administered to FBJ cells with different metastatic capacities. Production of glycosylated products derived from Xyl-Ser-C12, especially heparan sulfate (HS) GAG-type oligosaccharides such as GalNAc-GlcA-GlcNAc-GlcA-Gal-Gal-Xyl-Ser-C12, was indicated in poorly metastatic FBJ-S1 cells more than in highly metastatic FBJ-LL cells by LC-MS. The results of RT-PCR revealed that HS synthases, Ext1 and Ext2, were expressed in FBJ-S1 cells more than in FBJ-LL cells. Furthermore, siRNA against Ext1 suppressed the expression of HS and enhanced the motility of FBJ-S1 cells. In addition, the expression of heparanase (HPSE) was enhanced in Ext-1-knockdown FBJ-S1 cells, and responsible for the increase in cell motility caused by the down-regulation of Ext1 expression. Our data provide the first evidence that Ext1 regulates the expression of HPSE and also indicated that levels of Ext1 and HPSE influenced the motility of FBJ cells.

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