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KMID : 0603520000050020071
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Journal of Korean Association of Cancer Prevention
2000 Volume.5 No. 2 p.71 ~ p.80
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Anti-angiogenic Effect of Tissue Inhibitor of Metalloproteinase-3 (TIMP-3)from shark, Scyliorhinus torazame
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Kim Ji-Tae
Jung Joo-Won
Sohn Tae-Kwon
Kim Kyu-Won
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Abstract
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Shark cartilage has been reported to strongly inhibit the primary tumor, however, anti-angiogenic substances from this tissue were not elucidated yet. Previously, we cloned the full-length cDNA encoding TIMP-3 known to inhibit the angiogenesis of primary tumor, from the cartilage of Scyliorhinus torazame. In order to investigate the anti-angiogenic activity of shark TIMP-3 (sTIMP-3), we initially constructed sTIMP-3 transfected HT1080 human fibrosarcoma cells. In vitro invasion and migration assays showed that concentrated conditioned media (CCM) of sTIMP-3 transfectant decreased the invasive activity and migration of human umbilical vein endothelial cells (HUVECs). Also, in in vitro tube formation assay, CCM of sTIMP-3 transfected HT1080 decreased the ability of tube formation in HUVECs. To determine whether sTIMP-3 had an effect of anti-angiogenesis in vivo, we used chorioallantoic membrane (CAM) assay with CCM of NIH3T3 transfectant. CCM of sTIMP-3 transfected NIH3T3 inhibited new blood vessels of chicken embryo. In addition, to produce recombinant sTIMP-3 protein, prokaryotic TIMP-3 expression vector was constructed and induced. IN CAM assay on recombinant sTIMP-3 fusion protein produced in E. coli, formation of new blood vessels was inhibited too. These results suggest that TIMP-3 of Scyliorhinus torazame has an anti-angiogenic activity.
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KEYWORD
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Shark, TIMP, Angiogenesis
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