Activation of calcium-dependent kinases and epidermal growth factor receptor regulate muscarinic acetylcholine receptor-mediated MAPK/ERK activation in thyroid epithelial cells
Activation of calcium-dependent kinases and epidermal growth factor receptor regulate muscarinic acetylcholine receptor-mediated MAPK/ERK activation in thyroid epithelial cells
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DOI：
10.1016/j.cellsig.2007.06.010
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发表时间：
2007-10-01
影响因子：
4.8
通讯作者：
Jimenez, Eugenio
中科院分区：
文献类型：
--
作者：
Montiel, Mercedes;Quesada, Juan;Jimenez, Eugenio
我们以前表明，毒蕈碱乙酰胆碱受体（mAChR）的刺激卡巴胆碱（Cch）引起的时间和剂量依赖性增加的丝裂原活化蛋白激酶/细胞外信号调节激酶（MAPK/ERK）的磷酸化在甲状腺上皮细胞。在这项研究中，我们证明，mAChR刺激也诱导了时间依赖性增加的酪氨酸磷酸化的脯氨酸丰富的酪氨酸激酶2（Pyk 2），这是防止预处理的甲状腺上皮细胞与特定的Src家族酪氨酸激酶抑制剂PP 2。此外，Pyk 2的磷酸化可通过螯合细胞外Ca ~（2+）或抑制磷脂酶C（PLC）而减弱，并可被一种特异性的微粒体Ca ~（2+）-ATP酶抑制剂毒胡萝卜素（thapsigargin）所诱导。在甲状腺上皮细胞中掺入Pyk 2反义寡核苷酸以下调Pyk 2表达或用Ca 2 +/钙调蛋白蛋白激酶II预处理细胞。(CaM激酶II）抑制剂KN-62显著降低Cch诱导的MAPK/ERK磷酸化。此外，磷脂酰肌醇3-激酶（PI 3 K）的选择性抑制剂LY 294002和渥曼青霉素、表皮生长因子受体（EGFR）激酶的特异性抑制剂tyrphostin AG 1478和小G蛋白异戊二烯化的翻译后抑制剂（-）-紫苏酸可部分抑制Cch诱导的MAPK/ERK磷酸化。综上所述，我们的数据表明，Pyk 2，CaM激酶11和Src家族酪氨酸激酶是通过EGFR/Ras/Raf通路激活MAPK/ERK级联反应的关键分子在甲状腺上皮细胞中响应于mAChR刺激。(c)2007年爱思唯尔公司All rights reserved.
We previously showed that stimulation of muscarinic acetylcholine receptors (mAChR) by carbachol (Cch) caused a time- and dose-dependent increase of mitogen-activated protein kinase/extracellular signal-regulated kinases (MAPK/ERK) phosphorylation in thyroid epithelial cells. In this study, we demonstrated that mAChR stimulation also induced a time-dependent increase in the tyrosine phosphorylation of proline-rich tyrosine kinase 2 (Pyk2), which was prevented by pretreatment of thyroid epithelial cells with the specific Src-family tyrosine kinase inhibitor PP2. Besides, phosphorylation of Pyk2 was attenuated by chelation of extracellular Ca2+ or inhibition of phospholipase C (PLC), and was evoked by thapsigargin, a specific microsomal Ca2+-ATPase inhibitor. Incorporation of Pyk2 antisense oligonucleotides in thyroid epithelial cells to downregulated Pyk2 expression or pretreatment of cells with the Ca2+/calmodulin protein kinase II. (CaM kinase II) inhibitor KN-62 significantly reduced Cch-induced MAPK/ERK phosphorylation. In addition, Cch-induced MAPK/ERK phosphorylation was partially inhibited by LY294002 and wortmannin, two selective inhibitors of phosphatidylinositol 3-kinase (PI3K), tyrphostin AG1478, a specific inhibitor of epidermal growth factor receptor (EGFR) kinase, and (-)-perillic acid, a post-translational inhibitor of small G-proteins isoprenylation. Taken together, our data suggest that Pyk2, CaM kinase 11 and Src-family tyrosine kinases are key molecules for the activation of MAPK/ERK cascade through the EGFR/Ras/Raf pathway in thyroid epithelial cells in response to mAChR stimulation. (c) 2007 Elsevier Inc. All rights reserved.