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
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
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DOI：
10.1038/labinvest.3780006
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发表时间：
2000-01-01
影响因子：
5
通讯作者：
Brown, CA
中科院分区：
文献类型：
--
作者：
Brown, J;Reading, SJ;Brown, CA
早期报告表明 ECV304 是源自日本人脐静脉内皮细胞 (HUVEC) 培养物的自发转化系。许多形态学、免疫化学和遗传学研究提供了进一步的证据，证明 ECV304 是一种有价值的生物医学研究工具，可用于研究包括体外血管生成和各种 G 蛋白偶联受体信号转导在内的过程。然而，ECV304 和 HUVEC 之间的几个明显差异现已显而易见，最近的报告表明 ECV304 和 T24/83（一种人类膀胱癌细胞系）之间的遗传相似性。为了进一步评估 ECV304 作为人内皮细胞模型的效用，我们比较了 ECV304 和 T24/83 对一系列 G 蛋白偶联受体激动剂的功能反应。我们还使用 DNA 指纹分析对 ECV304 和 T24/83 进行核型分析。 ATP 和三磷酸尿苷 (UTP) 均刺激 ECV304 中的磷酸肌醇代谢，但不改变 cAMP 水平。使用选择性 P2Y 受体激动剂的比较数据表明，这种导致细胞内储存的钙动员的反应主要是由 P2Y 受体的激活介导的。 ECV304 和 T24/83 细胞记录了类似的反应。 ECV304 表达相对较高的 NOS 基础活性，该活性可被 L-NAME 降低并被 P2Y 受体激动剂刺激。相反，P2Y受体激活并不诱导ECV304中前列腺素的合成。 ECV304 和 T24/83 均表达腺苷、肾上腺素和降钙素受体，刺激腺苷酸环化酶。 ECV304 和 T24/83 细胞的增殖（通过 [H-3] 胸苷掺入 DNA 来测量）在很大程度上不依赖于血清。这与猪和牛主动脉内皮细胞的平行实验形成鲜明对比，后者表明 DNA 合成显着依赖于血清。遗传分析证实ECV304和T24/83是相同的。 ECV304 显示一些内皮特征，可用于受体药理学研究。然而，ECV304 并非 HUVEC 来源，因此不适合研究内皮细胞生物学。
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, 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, receptor agonists. In contrast, P2Y, 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 [H-3]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.