Fibronectin- and protein kinase C-mediated activation of ERK/MAPK are essential for proplateletlike formation
Fibronectin- and protein kinase C-mediated activation of ERK/MAPK are essential for proplateletlike formation
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DOI：
10.1182/blood.v99.10.3579
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发表时间：
2002-05-15
期刊：
影响因子：
20.3
通讯作者：
Cohen, I
中科院分区：
文献类型：
--
作者：
Jiang, F;Jia, YZ;Cohen, I
巨核细胞CHRF-288细胞系用于研究负责前血小板样形成（PPF）的信号转导途径。纤维连接蛋白（FN）和蛋白激酶C（PKC）的激活在PPF中的作用进行了研究。在血清和佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA），PKC激活剂的存在下，细胞表现出完全的巨核细胞分化，表现为粘附，形状改变，细胞大小增加，多倍体，131317，和表达CD 41（+），CD 61（+），和CD 62 P（+）。在FN/PMA存在下，在无血清培养物中观察到相同的形态学和表型特征。只有部分分化发生时，其他整合素配体取代FN。FN单独诱导最小的细胞粘附和扩散，而PMA单独只诱导多倍体没有粘附。信号转导变化涉及细胞外信号调节蛋白激酶I（ERK 1）/ERK 2以及c-Jun氨基末端激酶1（JNK 1）/应激激活蛋白激酶（SAPK）的激活。磷酸肌醇-3激酶和p38在这些条件下没有刺激。抑制剂被用来确定信号通路和PPF之间的因果关系。PD 98059和GF 109203 X分别是ERK 1/ERK 2通路和PKC的抑制剂，可阻断PPF，而粘附、铺展和多倍体均正常。这些研究表明，ERK 1/ERK 2丝裂原活化蛋白激酶途径的激活在PPF中起着关键作用。阐明巨核细胞发育和PPF的信号转导通路对于理解这一独特的生物学过程至关重要。(C)2002年，美国血液学会。
The megakaryoblastic CHRF-288 cell line was used to investigate signal transduction pathways responsible for proplatelet-like formation (PPF). The role of fibronectin (FN) and protein kinase C (PKC) activation in PPF were examined. In the presence of serum and phorbol 12-myristate 13-acetate (PMA), a PKC activator, cells exhibited full megakaryocytic differentiation, manifested by adhesion, shape change, increased cell size, polyploidy, 131317, and expression of CD41(+), CD61(+), and CD62P(+). The same morphologic and phenotypic features were observed in serum-free cultures in the presence of FN/PMA. Only partial differentiation occurred when other integrin ligands were substituted for FN. FN alone induced minimal cell adhesion and spreading, while PMA alone induced only polyploidy without adhesion. Signal transduction changes involved the activation of the extracellular signal-regulated protein kinase I (ERK1)/ERK2 as well as c-Jun amino-terminal kinase 1 (JNK1)/stress-activated protein kinase (SAPK). Phosphoinositide-3 kinase and p38 were not stimulated under these conditions. Inhibitors were used to identify the causal relationship between signaling pathways and PPF. PD98059 and GF109203X, inhibitors of ERK1/ERK2 pathway and PKC, respectively, blocked PPF, while adhesion, spreading, and polyploidy were normal. These studies show that activation of ERK1/ERK2 mitogen-activated protein kinase pathway plays a critical role in PPF. The elucidation of the signal transduction pathway on megakaryocyte development and PPF is of crucial importance for understanding this unique biological process. (C) 2002 by The American Society of Hematology.