Critical Evaluation of ECV304 as a Human Endothelial Cell Model Defined by Genetic Analysis and Functional Responses: A Comparison with the Human Bladder Cancer Derived Epithelial Cell Line T24/83

Abstract

SUMMARY:Early reports indicated that ECV304 was a spontaneously-transformed line derived from a Japanese human umbilical vein endothelial cells (HUVEC) culture. Many morphological, immunochemical, and genetic studies provided further evidence that ECV304 was a valuable biomedical research tool and could be used to study processes that include angiogenesis in vitro and signal transduction by a variety of G protein-coupled receptors. However, several distinct differences between ECV304 and HUVEC are now apparent and recent reports have indicated genetic similarity between ECV304 and T24/83, a human bladder cancer cell line. To further assess the utility of ECV304 as a human endothelial cell model, we compared the functional responses of ECV304 and T24/83 to a range of G protein-coupled receptor agonists. We also used DNA fingerprinting to karyotype both ECV304 and T24/83. Both ATP and uridine triphosphate (UTP) stimulated inositol phosphate metabolism in ECV304 without alteration of cAMP levels. Comparative data using selective P2Y receptor agonists indicated that this response, leading to calcium mobilization from intracellular stores, was predominantly mediated by the activation of P2Y 2 receptors. Similar responses were recorded from both ECV304 and T24/83 cells. ECV304 expressed a relatively high basal activity of NOS that was reduced by L-NAME and stimulated by P2Y 2 receptor agonists. In contrast, P2Y 2 receptor activation did not induce prostaglandin synthesis in ECV304. Both ECV304 and T24/83 express receptors for adenosine, adrenaline, and calcitonin, which stimulate adenylate cyclase. Proliferation of ECV304 and T24/83 cells, measured by the incorporation of [ 3 H]thymidine into DNA, was largely serum-independent. This was in contrast to parallel experiments with porcine and bovine aortic endothelial cells that indicated a marked serum-dependent increase in DNA synthesis. Genetic analysis confirmed that ECV304 and T24/83 are identical. ECV304 displays some endothelial characteristics and is useful for the study of receptor pharmacology. However, ECV304 is not of HUVEC origin and is therefore an inappropriate cell line to study endothelial cell biology. (Lab Invest 2000, 80:37-45).

Atorvastatin prevents hypoxia-induced inhibition of endothelial nitric oxide synthase expression but does not affect heme oxygenase-1 in human microvascular endothelial cells

“…Moreover, in both studies the authors employed ECV304 cells, a cell line that has been considered as being of endothelial origin. However, recent studies convincingly demonstrated that they are derived from epithelial cells of bladder cancer [16].…”

Vaccination with xenogeneic tumor endothelial proteins isolated in situ inhibits tumor angiogenesis and spontaneous metastasis

“…Recent reports revealed genetic similarities between ECV304 and T24, a human bladder epithelial cancer cell line, by comparing their DNA fingerprints. 45,46 Nevertheless, ECV304 shows many endothelial characteristics, resembles Cl-2, a porcine brain microvascular endothelial cells and differs from T24 in phenotype. 47 ECV304 cells not only show endothelial morphology, but also express endothelial markers, uptake Ac-LDL and undergo tube formation in Matrigel.…”

Controlled light exposure microscopy reveals dynamic telomere microterritories throughout the cell cycle

“…ECV-304 was originally described as an immortalized endothelial cell line (32). However, more recent evidence has suggested that ECV-304 is a derivative of the human bladder carcinoma cell line T24 (33). For transfections and imaging, cells were grown on collagen coated glass bottom dishes (Matek).…”