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Primary ovarian carcinomas and abdominal metastasis contain 4,6-disulfated chondroitin sulfate rich regions, which provide adhesive properties to tumour cells.

Vallen MJ, Schmidt S, Oosterhof A, Bulten J, Massuger LF, van Kuppevelt TH - PLoS ONE (2014)

Bottom Line: Increased expression of CSE reduced cell migration.Addition of free CSE had similar effects.The data presented here indicate that CSE is associated with metastatic lesions and that it provides tumours with adhesive properties.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, The Netherlands; Department of Obstetrics and Gynaecology, Radboud University Medical Center, Nijmegen, The Netherlands.

ABSTRACT
High mortality in ovarian cancer patients is primarily caused through rapid metastasis of the tumour, but the underlying mechanisms are poorly understood. Glycosaminoglycans, are abundantly present in tumours and chondroitin sulfate-E (CSE), a highly 4,6-sulfated glycosaminoglycan, has been indicated to play a role in carcinogenesis. In this study we investigated the presence of CSE in ovarian cancer metastasis and studied its role in tumour cell adhesiveness and migration. CSE was studied immunohistochemically in primary ovarian carcinomas and abdominal metastases using the single chain antibody GD3G7. The role of CSE was studied in 2D (scratch assays) and 3D (collagen matrices, spheroids) systems using SKOV3 cells applying 1: overexpression of CSE by stable transfection with DNA encoding GalNAc4S-6 sulfotransferase, 2: enzymatic removal of CS, and 3: addition of CSE. In ovarian cancer tissue, CSE expression was predominantly seen in the stromal compartment of both primary ovarian carcinomas and metastases, with a comparable degree of intensity and extent. Overexpression of CSE disaccharide units by tumour cells increased their adhesive properties which was especially seen in tumour spheroid formation. Increased expression of CSE reduced cell migration. Addition of free CSE had similar effects. The data presented here indicate that CSE is associated with metastatic lesions and that it provides tumours with adhesive properties. CSE rich motifs are put forward as a potential target for ovarian cancer therapy.

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CSE inhibits SKOV3 cell migration out of spheroids in 3D collagen matrices.A) Wildtype and cells transfected with an empty vector (-) showed cell migration out of spheroids, whereas migration of cells transfected with GalNAc4S-6STwas almost absent. B) Strong restriction of migration of wildtype SKOV3 cells out of spheroids in CSE decorated collagen matrices. Bar: 200 µm.
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pone-0111806-g007: CSE inhibits SKOV3 cell migration out of spheroids in 3D collagen matrices.A) Wildtype and cells transfected with an empty vector (-) showed cell migration out of spheroids, whereas migration of cells transfected with GalNAc4S-6STwas almost absent. B) Strong restriction of migration of wildtype SKOV3 cells out of spheroids in CSE decorated collagen matrices. Bar: 200 µm.

Mentions: To study the effect of CSE on cell migration in 3D systems, spheroids were prepared from wildtype and transfected SKOV3 cells and embedded in a collagenous matrix. Wildtype SKOV3 cells migrated out of the spheroid primarily as single cells and sometimes following collective strand formation. The same was seen for spheroids prepared from SKOV3 cells harboring an empty vector. However, SKOV3 cells overexpressing CSE were hardly able to migrate out of the spheroid, nor as single cells, nor as collective strands (fig. 7A). In an alternative approach, we prepared collagenous matrices in the presence of CSE, resulting in collagen fibrils associated with CSE. In such matrices wildtype SKOV3 cells showed very restricted cell migration (fig. 7B), further indicating the cell migrative inhibiting effect of CSE.


Primary ovarian carcinomas and abdominal metastasis contain 4,6-disulfated chondroitin sulfate rich regions, which provide adhesive properties to tumour cells.

Vallen MJ, Schmidt S, Oosterhof A, Bulten J, Massuger LF, van Kuppevelt TH - PLoS ONE (2014)

CSE inhibits SKOV3 cell migration out of spheroids in 3D collagen matrices.A) Wildtype and cells transfected with an empty vector (-) showed cell migration out of spheroids, whereas migration of cells transfected with GalNAc4S-6STwas almost absent. B) Strong restriction of migration of wildtype SKOV3 cells out of spheroids in CSE decorated collagen matrices. Bar: 200 µm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4221137&req=5

pone-0111806-g007: CSE inhibits SKOV3 cell migration out of spheroids in 3D collagen matrices.A) Wildtype and cells transfected with an empty vector (-) showed cell migration out of spheroids, whereas migration of cells transfected with GalNAc4S-6STwas almost absent. B) Strong restriction of migration of wildtype SKOV3 cells out of spheroids in CSE decorated collagen matrices. Bar: 200 µm.
Mentions: To study the effect of CSE on cell migration in 3D systems, spheroids were prepared from wildtype and transfected SKOV3 cells and embedded in a collagenous matrix. Wildtype SKOV3 cells migrated out of the spheroid primarily as single cells and sometimes following collective strand formation. The same was seen for spheroids prepared from SKOV3 cells harboring an empty vector. However, SKOV3 cells overexpressing CSE were hardly able to migrate out of the spheroid, nor as single cells, nor as collective strands (fig. 7A). In an alternative approach, we prepared collagenous matrices in the presence of CSE, resulting in collagen fibrils associated with CSE. In such matrices wildtype SKOV3 cells showed very restricted cell migration (fig. 7B), further indicating the cell migrative inhibiting effect of CSE.

Bottom Line: Increased expression of CSE reduced cell migration.Addition of free CSE had similar effects.The data presented here indicate that CSE is associated with metastatic lesions and that it provides tumours with adhesive properties.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, The Netherlands; Department of Obstetrics and Gynaecology, Radboud University Medical Center, Nijmegen, The Netherlands.

ABSTRACT
High mortality in ovarian cancer patients is primarily caused through rapid metastasis of the tumour, but the underlying mechanisms are poorly understood. Glycosaminoglycans, are abundantly present in tumours and chondroitin sulfate-E (CSE), a highly 4,6-sulfated glycosaminoglycan, has been indicated to play a role in carcinogenesis. In this study we investigated the presence of CSE in ovarian cancer metastasis and studied its role in tumour cell adhesiveness and migration. CSE was studied immunohistochemically in primary ovarian carcinomas and abdominal metastases using the single chain antibody GD3G7. The role of CSE was studied in 2D (scratch assays) and 3D (collagen matrices, spheroids) systems using SKOV3 cells applying 1: overexpression of CSE by stable transfection with DNA encoding GalNAc4S-6 sulfotransferase, 2: enzymatic removal of CS, and 3: addition of CSE. In ovarian cancer tissue, CSE expression was predominantly seen in the stromal compartment of both primary ovarian carcinomas and metastases, with a comparable degree of intensity and extent. Overexpression of CSE disaccharide units by tumour cells increased their adhesive properties which was especially seen in tumour spheroid formation. Increased expression of CSE reduced cell migration. Addition of free CSE had similar effects. The data presented here indicate that CSE is associated with metastatic lesions and that it provides tumours with adhesive properties. CSE rich motifs are put forward as a potential target for ovarian cancer therapy.

Show MeSH
Related in: MedlinePlus