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Background:Tumour stromal cells differ from its normal counterpart. We have shown that tumour endothelial cells (TECs) isolated from tumour tissues are also abnormal. Furthermore, we found that mRNAs of vascular endothelial growth factor-A (VEGF-A) and cyclooxygenase-2 (COX-2) were upregulated in TECs. Vascular endothelial growth factor-A and COX-2 are angiogenic factors and their mRNAs contain an AU-rich element (ARE). AU-rich element-containing mRNAs are reportedly stabilised by Hu antigen R (HuR), which is exported to the cytoplasm.Methods:Normal endothelial cell (NEC) and two types of TECs were isolated. We evaluated the correlation of HuR and accumulation of VEGF-A and COX-2 mRNAs in TECs and effects of HuR on biological phenotypes of TECs.Results:The HuR protein was accumulated in the cytoplasm of TECs, but not in NECs. Vascular endothelial growth factor-A and COX-2 mRNA levels decreased due to HuR knockdown and it was shown that these ARE-mRNA were bound to HuR in TECs. Furthermore, HuR knockdown inhibited cell survival, random motility, tube formation, and Akt phosphorylation in TECs.Conclusion:Hu antigen R is associated with the upregulation of VEGF-A and COX-2 mRNA in TECs, and has an important role in keeping an angiogenic switch on, through activating angiogenic phenotype in tumour endothelium.

Tumor blood vessels play an important role in tumor progression and metastasis. Thus, targeting tumor blood vessels is an important strategy for cancer therapy, especially for head and neck cancer patients. Tumor blood vessels generally sprout from pre‐existing vessels and have been thought to be genetically normal. However, tumor blood vessels have been shown to differ from their normal counterparts, for example, by changes in morphology. The authors isolated tumor endothelial cells (TECs) from mouse tumor xenografts and have shown that the TECs are abnormal. TECs up‐regulate many genes and proliferate more rapidly and migrate more than normal endothelial cells (NECs). Furthermore, TECs were found to be cytogenetically abnormal. We conclude that TECs can acquire cytogenetic abnormalities while in a tumor microenvironment. To develop ideal antiangiogenic therapies, understanding the crosstalk between blood vessels and the tumor microenvironment is important. Here, we provide an overview of the current studies on TEC abnormalities and a discussion about possible mechanisms for how tumor the microenvironment makes TECs abnormal.

An important concept in tumor angiogenesis has been that tumor blood vessels contain genetically normal and stable endothelial cells (ECs), unlike tumor cells, which typically display genetic instability. Chromosomal aberration in human tumor endothelial cells (hTECs) in carcinoma has not yet been investigated. Here we isolated TECs from 20 human renal cell carcinomas (RCCs) and analyzed their cytogenetic abnormalities. The degree of aneuploidy was analyzed by fluorescence in situ hybridization using chromosome 7 and chromosome 8 DNA probes in isolated hTECs. In human RCCs, 22-58% (median 33%) of uncultured hTECs were aneuploid, whereas normal ECs were diploid. The mechanisms of TEC aneuploidy were studied using mouse TECs (mTECs) isolated from xenografts of human epithelial tumors. To investigate the contribution of progenitor cells to aneuploidy in mTECs, CD133 positive and CD133 negative mTECs were compared for aneuploidy. CD133 positive mTECs showed aneuploidy more frequently than CD133 negative mTECs. This is the first report showing cytogenetic abnormality of hTECs in carcinoma, contrary to traditional belief. It is suggested that cytogenetic alterations in tumor vessels of carcinoma can occur and may play a signicant role in modifying tumor-stromal interactions. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1308.

A green tea polyphenol, epigallocatechin-3 gallate (EGCG) suppresses tumor growth in vivo. We investigated mechanisms of specific tumor angiogenesis inhibition using tumor-associated endothelial cells (TECs), peripheral blood-derived ECs, and normal endothelial cells (NECs). TECs were isolated and cultured from human tumor xenografts in nude mice. Peripheral blood-derived ECs were isolated from peripheral blood of nude mice. EGCG suppressed migration of TECs and peripheral blood-derived ECs. EGCG also inhibited the phosphorylation of Akt in TECs and peripheral blood-derived ECs. The PI3K inhibitor, LY294002 blocked the migration of TECs induced by VEGF. Furthermore, VEGF-induced mobilization of CD133/VEGFR-2 double-positive cells into circulation was inhibited by EGCG. MMP-9 in the bone marrow is involved in mobilizing bone marrow-derived VEGFR-2 positive cells into peripheral circulation. Expression of MMP-9 mRNA was suppressed in bone marrow stromal cells by EGCG. In vivo model, EGCG reduced melanoma growth. Our study showed that EGCG specifically inhibits TEC and circulating EC through PI3K in EC and MMP-9 expression in bone marrow stroma. EGCG is a promising angiogenesis inhibitor for cancer therapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3432.