β-adrenergic receptor activation inhibits keratinocyte migration via a cyclic adenosine monophosphate-independent mechanism
β-adrenergic receptor activation inhibits keratinocyte migration via a cyclic adenosine monophosphate-independent mechanism
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DOI：
10.1046/j.1523-1747.2002.19611.x
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发表时间：
2002-12-01
影响因子：
6.5
通讯作者：
Isseroff, RR
中科院分区：
文献类型：
--
作者：
Chen, J;Hoffman, BB;Isseroff, RR
越来越多的证据表明，在多种细胞类型中，g蛋白偶联受体与生长因子受体介导的信号转导相互作用。我们研究了β -肾上腺素能受体（典型的gtp结合蛋白偶联受体）的激活影响培养的人角质形成细胞迁移的机制。我们发现异丙肾上腺素，一种β -肾上腺素能受体选择性激动剂，以浓度依赖的方式抑制表皮生长因子或细胞外Ca2+刺激的细胞迁移。这可以通过β -肾上腺素能受体选择性拮抗剂替莫洛尔对细胞进行预处理来预防。有趣的是，异丙肾上腺素在浓度为1 nM时，并没有明显增加细胞内环磷酸腺苷的浓度，但却抑制了50%的细胞迁移。为了进一步测试异丙肾上腺素的作用是否通过腺苷酸环化酶的激活介导，使用了其活性的两种抑制剂，2'5'-二脱氧腺苷和SQ22536。这两种化合物显著降低异丙肾上腺素诱导的细胞内环磷酸腺苷浓度的增加，但没有减弱异丙肾上腺素诱导的细胞迁移抑制。此外，福斯克林（1mum）显著增加细胞内环磷酸腺苷浓度，但不显著抑制细胞迁移。由于已知有丝分裂原激活的蛋白激酶是生长因子刺激的细胞迁移的信号，我们研究了β -肾上腺素能受体介导的角质形成细胞迁移的抑制是否可能通过有丝分裂原激活的蛋白激酶的失活而发生。我们发现异丙肾上腺素以浓度依赖性的方式抑制细胞外信号调节激酶丝裂原活化蛋白激酶的磷酸化，但对应激丝裂原活化蛋白激酶c-jun n-末端激酶和应激活化蛋白激酶-2的磷酸化没有影响。无论是福斯克林还是一种膜渗透性环腺苷单磷酸类似物都不能抑制这些有丝分裂原激活的蛋白激酶的磷酸化。这些发现表明，β -肾上腺素能受体诱导的角质形成细胞迁移的抑制是通过抑制细胞外信号调节的激酶丝裂原激活的蛋白激酶信号传导以不依赖环腺苷单磷酸的方式介导的。
There is increasing evidence that G-protein-coupled receptors cross-talk with growth factor receptor-mediated signal transduction in a variety of cell types. We have investigated mechanisms by which the activation of beta-adrenergic receptors, classically GTP-binding proteins coupled receptors, influence the migration of cultured human keratinocytes. We found that iso-proterenol, a beta-adrenergic receptor-selective agonist, inhibited cell migration stimulated by either epidermal growth factor, or extracellular Ca2+ in a concentration-dependent manner. This was prevented by pretreatment of the cells with the beta-adrenergic receptor-selective antagonist timolol. Interestingly, isoproterenol, at a concentration of 1 nM, did not measurably increase intracellular cyclic adenosine monophosphate concentrations yet inhibited cell migration by 50%. To test further if isoproterenol's actions were mediated via activation of adenylyl cyclase, two inhibitors of its activity, 2'5'-dideoxyadenosine and SQ22536, were used. Both compounds significantly diminished iso-proterenol-induced increases in intracellular cyclic adenosine monophosphate concentrations but did not attenuate isoproterenol-induced inhibition of cell migration. Also, forskolin (1 muM) markedly increased intracellular cyclic adenosine monophosphate concentrations but did not significantly inhibit cell migration. As mitogen-activated protein kinases are known to signal growth factor-stimulated cell migration, we examined whether beta-adrenergic receptor-mediated inhibition of keratinocyte migration might occur via inactivation of mitogen-activated protein kinases. We found that isoproterenol inhibited phosphorylation of extracellular signal-regulated kinase mitogen-activated protein kinase in a concentration-dependent manner but had no effect on the phosphorylation of the stress mitogen-activated protein kinases c-jun N-terminal kinase and stress-activated protein kinase-2. Neither forskolin nor a membrane permeable cyclic adenosine monophosphate analog inhibited phosphorylation of any of these mitogen-activated protein kinases. These findings suggest that beta-adrenergic receptor-induced inhibition of keratinocyte migration is mediated through inhibition of the extracellular signal-regulated kinase mitogen-activated protein kinase signaling in a cyclic adenosine monophosphate-independent manner.