Journal
BRITISH JOURNAL OF CANCER
Volume 88, Issue 9, Pages 1453-1461Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/sj.bjc.6600907
Keywords
angiogenesis; IL-12; VEGFR-3; image processing
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Funding
- NCI NIH HHS [CA28332, R01 CA028332, R01 CA052586, CA52586] Funding Source: Medline
- NIAID NIH HHS [T32AI07285, T32 AI007285] Funding Source: Medline
- NIGMS NIH HHS [GM67143, R01 GM067143] Funding Source: Medline
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New blood vessel formation within tumour's is a critical feature for tumour growth. A major limitation in understanding this complex process has been the inability to visualise and analyse vessel formation. Here, we report on the development of a whole-tissue mount technique that allows visualisation of vessel structure. Mice expressing green fluorescent protein (GFP) made it possible to easily see GFP(+) vessels within non-GFP-expressing B 16 melanoma tumours. The small fragments of tumour used in this technique were also effectively stained with fluorescent probe-conjugated antibodies, allowing characterisation of the vessels based on surface marker phenotype. The vessels within tumour tissue were much more irregular and tortuous compared to those within surrounding normal muscle. B 16 tumours stably transfected with the genes for IL-12 were used to assess the effects of this cytokine on tumour growth and vessel formation. The IL-12-expressing tumours grew more slowly and had much smaller blood vessels than the large, webbed vessels characteristic of the parental tumours, effects that were dependent on interferon gamma (IFN-gamma). Vessels in the parental tumours were found to express VEGFR-3, the receptor for VEGF-C and VEGF-D. Expression of this receptor by the endothelial cells of the blood vessels was lost in the cytokine expressing tumours, thus suggesting a mechanism for the antiangiogenic effects of IL-12. The combination of the whole mount technique and the GFP transgenic mice provides a powerful method for visualising tumour vasculature and characterising the effects of agents such as cytokines. (C) 2003 Cancer Research UK.
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