Identification of adenosine receptor subtypes expressed in the human endothelial-like ECV304 cells
Identification of adenosine receptor subtypes expressed in the human endothelial-like ECV304 cells
复制标题
DOI：
10.1159/000084547
复制
发表时间：
2005-01-01
期刊：
影响因子：
3.1
通讯作者：
Nakata, H
中科院分区：
文献类型：
--
作者：
Kobayashi, R;Saitoh, O;Nakata, H
腺苷通过其受体起作用，是内皮细胞的有效内源性调节剂。因此，培养表达腺苷受体的内皮细胞对于阐明腺苷在这些细胞系统中的功能的分子机制是重要的。因此，在来源于人脐静脉内皮细胞培养物的人ECV 304细胞系中进行腺苷受体的鉴定。RT-PCR实验显示，ECV 304细胞表达A（1）和A（2B）腺苷受体的mRNA。A（2A）腺苷受体mRNA表达水平不明显，A（3）腺苷受体mRNA表达水平不明显。使用各种放射性标记的腺苷受体配体对ECV 304细胞膜组分进行的结合研究表明，存在A（1）腺苷受体245 fmol/ mg膜蛋白，但发现A（2A）和A（3）腺苷受体的特异性结合可忽略不计。通过腺苷-3 '，5'-环磷酸和细胞外信号调节激酶的测定，检测了A（1）和A（2B）腺苷受体在ECV 304细胞中的功能表达，但在所采用的测定条件下，A（2A）腺苷受体的功能表达未得到证实。本研究结果表明，人内皮样细胞ECV 304中存在功能性腺苷A（1）和A（2B）受体，提示人内皮样细胞ECV 304可作为研究腺苷受体，特别是腺苷A2 B受体的良好模型。版权所有（C）2005 S. Karger AG，巴塞尔。
Adenosine, acting through its receptors, is a potent endogenous regulator of endothelial cells. The cultured endothelial cells expressing adenosine receptors are thus important for elucidation of molecular mechanism of adenosine functions in these cell systems. Therefore, identification of adenosine receptors in the human ECV304 cell line derived from a human umbilical vein endothelial cell culture was performed. RT-PCR experiments revealed that ECV304 cells express mRNAs for A(1) and A(2B) adenosine receptors. The expression of mRNA for A(2A) adenosine receptor was not in a significant level and that for A(3) adenosine receptor was not detected. The binding study of ECV304 cell membrane fractions using various radiolabeled ligands for adenosine receptors indicated the presence of A(1) adenosine receptors 245 fmol/ mg of membrane proteins, but the specific binding for A(2A) and for A(3) adenosine receptors were found to be negligible. The functional expression of A(1) and A(2B) adenosine receptors in ECV304 cells was detected by assays for adenosine- 3', 5'-cyclic monophosphate and for extracellular signal-regulated kinase, but that of A(2A) adenosine receptors was not confirmed under the assay conditions employed. In conclusion, this study presented evidence for functional A(1) and A(2B) adenosine receptors in human endothelial-like ECV304 cells, indicating that ECV304 cells can be a good model for the study of adenosine receptors, especially for A2B adenosine receptor, in endothelial cells. Copyright (C) 2005 S. Karger AG, Basel.