Busto R, Carrero I, Zapata P, Colás B, Prieto J C
Department of Biochemistry and Molecular Biology, University of Alcalá, E-28871, Alcalá de Henares, Spain.
Regul Pept. 2000 Nov 24;95(1-3):53-8. doi: 10.1016/s0167-0115(00)00129-4.
The pharmacological profile of adenylyl cyclase activity was analysed in WI-38 human foetal lung fibroblasts. Among various agents that act through G-protein coupled receptors, only the beta-adrenergic agonist isoproterenol stimulated and the tetradecapeptide somatostatin (SRIF, sst) inhibited the enzyme activity. The use of the reverse transcription-polymerase chain reaction (RT-PCR) methodology with appropriate cDNAs allowed us to identify the expression of four subtypes of SRIF transmembrane receptors (sst1-4 but not sst5 receptors) in this cell line. By RT-PCR and immunochemistry techniques, we also demonstrated the expression of stimulatory (alpha(s)) and inhibitory (alpha(i1), alpha(i2) and alpha(i3)) G-protein subunits. The known role of the adenylyl cyclase system in cell proliferation and differentiation mechanisms together with the present analysis of the corresponding regulatory network in fibroblasts of human foetal lung add knowledge on the cell line WI-38 that is widely used as a model system in studying cell growth. The importance of this cell class in normal and abnormal lung function and development reinforces the significance of these results.
在WI-38人胚肺成纤维细胞中分析了腺苷酸环化酶活性的药理学特征。在通过G蛋白偶联受体起作用的各种试剂中，只有β-肾上腺素能激动剂异丙肾上腺素刺激该酶活性，而十四肽生长抑素（SRIF，sst）抑制该酶活性。使用逆转录聚合酶链反应（RT-PCR）方法和合适的cDNA使我们能够鉴定出该细胞系中四种SRIF跨膜受体亚型（sst1-4，但不是sst5受体）的表达。通过RT-PCR和免疫化学技术，我们还证明了刺激性（α(s)）和抑制性（α(i1)、α(i2)和α(i3)）G蛋白亚基的表达。腺苷酸环化酶系统在细胞增殖和分化机制中的已知作用，以及目前对人胚肺成纤维细胞中相应调节网络的分析，增加了对WI-38细胞系的了解，该细胞系在研究细胞生长中被广泛用作模型系统。这类细胞在正常和异常肺功能及发育中的重要性强化了这些结果的意义。
