Activation of MAP kinase by muscarinic cholinergic receptors induces cell proliferation and protein synthesis in human breast cancer cells
Activation of MAP kinase by muscarinic cholinergic receptors induces cell proliferation and protein synthesis in human breast cancer cells
复制标题
DOI：
10.1002/jcp.20326
复制
发表时间：
2005-08-01
影响因子：
5.6
通讯作者：
Montiel, M
中科院分区：
文献类型：
--
作者：
Jiménez, E;Montiel, M
卡巴胆碱（Carbachol，Cch）是一种毒蕈碱型乙酰胆碱受体（mAChR）激动剂，可增加MCF-7人乳腺癌细胞内游离Ca（2+）的动员，并诱导丝裂原活化蛋白激酶/细胞外信号调节激酶（MAPK/ERK）磷酸化。用选择性磷脂酶C（PLC）抑制剂U 73122预处理细胞，或在无Ca 2+培养基中孵育细胞并不改变Cch刺激的MAPK/ERK磷酸化。MAPK/ERK的磷酸化被蛋白激酶C（PKC）的激活剂佛波酯12-肉豆蔻酸酯乙酸酯（PMA）模拟，但Cch诱导的MAPK/ERK激活不受PKC下调或用PKC抑制剂GF 109203 X预处理细胞的影响。然而，Cch刺激的MAPK/ERK磷酸化完全阻断豆蔻酰化PKC-xi假底物，PKC-xi的特异性抑制剂，和高剂量的星形孢菌素。用磷脂酰肌醇3-激酶（PI 3 K）的选择性抑制剂wortmannin或LY 294002预处理人乳腺癌细胞，可减少Cch介导的MAPK/ERK磷酸化。当MCF-7细胞用染料木黄酮（一种非选择性酪氨酸激酶抑制剂）或特异性Src酪氨酸激酶抑制剂PP 2预处理时，观察到类似的结果。此外，在MCF-7人乳腺癌细胞中，mAChR刺激诱导蛋白质合成和细胞增殖增加，并且这些作用被有丝分裂原活化激酶激酶的特异性抑制剂PD 098059阻止。总之，对mAChR下游效应物的分析表明，PKC-xi、PI 3 K和Src酪氨酸激酶家族是导致MAPK/ERK激活的信号级联中的关键分子，而不是细胞内游离Ca 2+动员或常规和新型PKC激活。此外，MAPK/ERK参与MCF-7人乳腺癌细胞生长和增殖的调节。
Carbachol (Cch), a muscarinic acetylcholine receptor(mAChR) agonist, increases intracellular-freeCa(2+) mobilization and induces mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) phosphorylation in MCF-7 human breast cancer cells. Pretreatment of cells with the selective phospholipase C (PLC) inhibitor U73122, or incubation of cells in a Ca2+-free medium did not alter Cch-stimulated MAPK/ERK phosphorylation. Phosphorylation of MAPK/ERK was mimicked by phorbol 12-myristate acetate (PMA), an activator of protein kinase C (PKC), but Cch-evoked MAPK/ERK activation was unaffected by down-regulation of PKC or by pretreatment of cells with GF109203X, a PKC inhibitor. However, Cch-stimulated MAPK/ERK phosphorylation was completely blocked by myristoylated PKC-xi pseudosubstrate, a specific inhibitor of PKC-xi, and high doses of staurosporine. Pretreatment of human breast cancer cells with wortmannin or LY294002, selective inhibitors of phosphoinositide 3-kinase (PI3K), diminished Cch-mediated MAPK/ERK phosphorylation. Similar results were observed when MCF-7 cells were pretreated with genistein, a non-selective inhibitor of tyrosine kinases, or with the specific Src tyrosine kinase inhibitor PP2. Moreover, in MCF-7 human breast cancer cells mAChR stimulation induced an increase of protein synthesis and cell proliferation, and these effects were prevented by PD098059, a specific inhibitor of the mitogen activated kinase kinase. In conclusion, analyses of mAChR downstream effectors reveal that PKC-xi, PI3K, and Src family of tyrosine kinases, but not intracellular-free Ca2+ mobilization or conventional and novel PKC activation, are key molecules in the signal cascade leading to MAPK/ERK activation. In addition, MAPK/ERK are involved in the regulation of growth and proliferation of MCF-7 human breast cancer cells.