Miggin S M, Kinsella B T
Department of Biochemistry, Merville House, University College Dublin, Belfield, Dublin 4, Ireland.
Biochim Biophys Acta. 1998 Nov 27;1425(3):543-59. doi: 10.1016/s0304-4165(98)00109-3.
The human thromboxane A2 receptor (TP), a G protein-coupled receptor, exists as two isoforms, TPalpha and TPbeta, which arise by alternative mRNA splicing and differ exclusively in their carboxyl terminal cytoplasmic regions. In this study, a reverse transcriptase-polymerase chain reaction (RT-PCR)-based strategy was developed to examine the expression of the TPs in tissues of physiologic relevance to TXA2. Although most of the 17 different cell/tissue types examined expressed both TP isoforms, the liver hepatoblastoma HepG2 cell line was found to exclusively express TPalpha mRNA. In most cell types, TPalpha mRNA predominated over TPbeta mRNA. Moreover, although the levels of TPalpha mRNA expression were similar in most of the cell/tissue types examined, extensive differences in the levels of TPbeta mRNA were observed. Consequently, the relative expression of TPalpha: TPbeta mRNA varied considerably due to extensive differences in TPbeta mRNA expression. Most strikingly, primary HUVECs were found to express: (i) low levels of TPbeta and (ii) approximately 6-fold greater levels of TPalpha than TPbeta. These data were confirmed in the spontaneously transformed HUVEC derived ECV304 cell line. Expression of TP mRNAs in the various tissue/cells correlated with protein expression, as assessed by radioligand binding using the selective TP antagonist [3H]SQ29,548.
人血栓素A2受体(TP)是一种G蛋白偶联受体,以两种异构体形式存在,即TPα和TPβ,它们通过选择性mRNA剪接产生,仅在其羧基末端胞质区域有所不同。在本研究中,开发了一种基于逆转录聚合酶链反应(RT-PCR)的策略,以检测与血栓素A2生理相关的组织中TP的表达。尽管所检测的17种不同细胞/组织类型中的大多数都表达两种TP异构体,但发现肝癌细胞系HepG2仅表达TPα mRNA。在大多数细胞类型中,TPα mRNA的表达量高于TPβ mRNA。此外,尽管在所检测的大多数细胞/组织类型中TPα mRNA的表达水平相似,但观察到TPβ mRNA水平存在广泛差异。因此,由于TPβ mRNA表达的广泛差异,TPα:TPβ mRNA的相对表达有很大变化。最显著的是,发现原代人脐静脉内皮细胞(HUVEC)表达:(i)低水平的TPβ,(ii)TPα水平约为TPβ的6倍。这些数据在自发转化的源自HUVEC的ECV304细胞系中得到证实。通过使用选择性TP拮抗剂[3H]SQ29548进行放射性配体结合评估,各种组织/细胞中TP mRNA的表达与蛋白质表达相关。