The role of TG2 in ECV304-related vasculogenic mimicry

Richard A. Jones, Zhuo Wang, Shakthi Dookie, Martin Griffin

Research output: Contribution to journalArticle

Abstract

Tumour vasculogenesis can occur by a process referred to as vasculogenic mimicry, whereby the vascular structures are derived from the tumour itself. These tumours are highly aggressive and do not respond well to anti-angiogenic therapy. Here, we use the well characterised ECV304 cell line, now known as the bladder cancer epithelial cell line T24/83 which shows both epithelial and endothelial characteristics, as a model of in vitro vasculogenic mimicry. Using optimised ratios of co-cultures of ECV304 and C378 human fibroblasts, tubular structures were identifiable after 8 days. The tubular structures showed high levels of TG2 antigen and TG in situ activity. Tubular structures and in situ activity could be blocked either by site-directed irreversible inhibitors of TG2 or by silencing the ECV304 TG2 by antisense transfection. In situ activity for TG2 showed co-localisation with both fibronectin and collagen IV. Deposition of these proteins into the extracellular matrix could be reduced by inclusion of non-cell penetrating TG inhibitors when analysed by Western blotting suggesting that the contribution of TG2 to tube formation is extracellular. Incubation of ECV304 cells with these same irreversible inhibitors reduced cell migration which paralleled a loss in focal adhesion assembly, actin cytoskeleton formation and fibronectin deposition. TG2 appears essential for ECV304 tube formation, thus representing a potential novel therapeutic target in the inhibition of vasculogenic mimicry. © 2012 Springer-Verlag.
LanguageEnglish
Pages89-101
Number of pages13
JournalAmino Acids
Volume44
Issue number1
Early online date10 Jan 2012
DOIs
Publication statusPublished - Jan 2013

Fingerprint

Tumors
Fibronectins
Cell Line
Neoplasms
Focal Adhesions
Extracellular Matrix Proteins
Fibroblasts
Coculture Techniques
Actin Cytoskeleton
Cell culture
Urinary Bladder Neoplasms
Cell Movement
Transfection
Blood Vessels
Actins
Collagen
Adhesion
Western Blotting
Epithelial Cells
Cells

Keywords

  • tissue transglutaminase
  • ECV304
  • co-culture
  • vasculogenic mimicry
  • inhibitors

Cite this

Jones, Richard A. ; Wang, Zhuo ; Dookie, Shakthi ; Griffin, Martin. / The role of TG2 in ECV304-related vasculogenic mimicry. In: Amino Acids. 2013 ; Vol. 44, No. 1. pp. 89-101.
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The role of TG2 in ECV304-related vasculogenic mimicry. / Jones, Richard A.; Wang, Zhuo; Dookie, Shakthi; Griffin, Martin.

In: Amino Acids, Vol. 44, No. 1, 01.2013, p. 89-101.

Research output: Contribution to journalArticle

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