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

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 diff...

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Bibliographic Details
Published in:Molecular and cellular biochemistry 2013-01, Vol.373 (1-2), p.63-72
Main Authors: Wang, Yinan, Yang, XiaoYan, Yamagata, Sadako, Yamagata, Tatsuya, Sato, Toshinori
Format: Article
Language:English
Subjects:
Animals
Biochemistry
Biomedical and Life Sciences
Biosynthesis
Carbohydrate Sequence
Cardiology
Cell adhesion & migration
Cell Line, Tumor
Cell migration
Cell Movement
Enzymes
Gene Expression
Gene Knockdown Techniques
Glucuronidase - genetics
Glucuronidase - metabolism
Glycosaminoglycans
Heparitin Sulfate - biosynthesis
Life Sciences
Medical Biochemistry
Mice
Molecular Sequence Data
N-Acetylglucosaminyltransferases - genetics
N-Acetylglucosaminyltransferases - metabolism
Oncology
Osteosarcoma
Physiological aspects
RNA, Small Interfering - genetics
Rodents
Sulfates
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Summary: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.
ISSN:0300-8177
1573-4919
DOI:10.1007/s11010-012-1475-8