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Carcinogenesis Advance Access published online on March 3, 2005
Carcinogenesis, doi:10.1093/carcin/bgi064
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© The Author 2005. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oupjournals.org
Received December 6, 2004
Revised February 21, 2005
Accepted February 22, 2005

CANCER BIOLOGY

Tumour regulation of fibroblast hyaluronan expression: a mechanism to facilitate tumour growth and invasion

M. Edward 1*, C. Gillan 1, D. Micha 1, and R. H. Tammi 2

1 Section of Squamous Cell Biology and Dermatology, Division of Cancer Sciences and Molecular Pathology, Robertson Building, University of Glasgow, Glasgow G12 8QQ, UK
2 Department of Anatomy, University of Kuopio, POB 1627, 70211 Kuopio, Finland

* To whom correspondence should be addressed.
M. Edward, E-mail: m.edward{at}clinmed.gla.ac.uk


  Abstract

Hyaluronan, a high molecular weight glycosaminoglycan is associated with cellular proliferation and migration. In a number of different tumour types, there is a close correlation between tumour progression and hyaluronan production, either by the tumour cells or the surrounding stromal cells. We have examined the ability of an aggressive melanoma cell line (C8161) to stimulate fibroblast hyaluronan synthesis, and the association of cell-surface CD44 receptors and hyaluronan with invasion.

Melanoma cell-conditioned medium prepared in low glucose medium (1 mg/ml) stimulated fibroblast glycosaminoglycan synthesis as measured by 3H-glucosamine incorporation, and hyaluronan synthesis as measured using a specific hyaluronan-binding plate assay, while tumour cell-conditioned medium prepared in high glucose medium (4.5 mg/ml) inhibited fibroblast glycosaminoglycan synthesis. High glucose tumour cell-conditioned medium contained large amounts of lactate that appeared to inhibit the tumour-derived factor stimulation of fibroblast glycosaminoglycan synthesis, as removal of the lactate restored the stimulating activity. Melanoma cells seeded on contracted collagen lattices and incubated at the air/liquid interface rapidly formed a multi-layered cell mass on the surface, with significant invasion of the gel. Hyaluronan staining was apparent within the collagen gel, and strong staining was seen around the invading tumour cells, but not around those cell layers near the surface. CD44 expression on the tumour cells was confined to those invading cells and corresponded to cellular hyaluronan staining. Hyaluronan staining was also apparent around and between tumour cells invading fibroblast-free collagen lattices. Monolayer cultures of C8161 cells stained strongly for CD44, but few cells stained for hyaluronan, while no detectable hyaluronan was released into the medium. In summary, the C8161 melanoma cells stimulated fibroblast hyaluronan synthesis, and in collagen lattices, only the invasive tumour cells expressed CD44 and hyaluronan, either in the presence or absence of fibroblasts.


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