Epidermal growth factor and thrombin induced proliferation of immortalized human keratinocytes is coupled to the synthesis of Egr-1, a zinc finger transcriptional regulator
Epidermal growth factor and thrombin induced proliferation of immortalized human keratinocytes is coupled to the synthesis of Egr-1, a zinc finger transcriptional regulator
复制标题
DOI：
10.1002/jcb.10145
复制
发表时间：
2002-01-01
影响因子：
4
通讯作者：
Thiel, G
中科院分区：
文献类型：
--
作者：
Kaufmann, K;Thiel, G
Western blotting显示，表皮生长因子（EGF）受体在HaCaT角质形成细胞中高表达。用EGF和丝氨酸蛋白酶凝血酶刺激HaCaT细胞，诱导DNA合成，通过将5-溴-2'-脱氧尿苷掺入增殖细胞的DNA来测量。利用针对EGF受体活性形式的抗体，我们发现在HaCaT细胞中，ECF和凝血酶触发了EGF受体的快速激活，随后是细胞外信号调节蛋白激酶（ERK）的磷酸化和激活。此外，EGF和凝血酶诱导锌指转录调节因子Egr-1的瞬时合成。MAP激酶抑制剂PD98059和EGF受体特异性酪氨酸激酶抑制剂AG1487完全抑制EGF或凝血酶处理的HaCaT细胞的增殖、ERK的激活和Egr-1的生物合成。这些数据表明，EGF受体和ERK的磷酸化和激活对于通过EGF和凝血酶进行的有丝分裂信号传导至关重要。EGF或凝血酶刺激导致HaCaT细胞中Egr-1的合成表明，Egr-1可能是EGF和凝血酶启动的信号级联反应的重要“晚期”部分。我们假设Egr-1可能作为角化细胞中的“第三信使”，将有丝分裂刺激与基因转录变化联系起来。j .细胞。生物化学学报，2002,31(2):381 -391。(C) 2002 Wiley-Liss, Inc。
The epidermal growth factor (EGF) receptor is highly expressed in HaCaT keratinocytes as shown by Western blotting. Stimulation of HaCaT cells with EGF, and also with the serine protease thrombin, induced DNA synthesis, measured by incorporation of 5-bromo-2'-deoxyuridine into the DNA of proliferating cells. Using antibodies directed against the active form of the EGF receptor, we show that in HaCaT cells ECF and thrombin triggered a rapid activation of the EGF receptor, followed by the phosphorylation and activation of the extracellular signal-regulated protein kinase (ERK). Moreover, EGF and thrombin induced a transient synthesis of the zinc finger transcriptional regulator Egr-1. Proliferation, activation of ERK, and biosynthesis of Egr-1 was completely inhibited in EGF or thrombin-treated HaCaT cells by the MAP kinase kinase inhibitor PD98059 and by AG1487, an EGF receptor-specific tyrosine kinase inhibitor. These data indicate that phosphorylation and activation of both the EGF receptor and ERK are essential for mitogenic signaling via EGF and thrombin. The synthesis of Egr-1 in HaCaT cells as a result of EGF or thrombin stimulation suggests that Egr-1 may be an important "late" part of the EGF and thrombin-initiated signaling cascades. We postulate that Egr-1 may function as a "third messenger" in keratinocytes connecting mitogenic stimulation with changes in gene transcription. J. Cell. Biochem. 85: 381 -391, 2002. (C) 2002 Wiley-Liss, Inc.