Requirement for PLCγ2 in IL-3 and GM-CSF-Stimulated MEK/ERK Phosphorylation in Murine and Human Hematopoietic Stem/Progenitor Cells
Requirement for PLCγ2 in IL-3 and GM-CSF-Stimulated MEK/ERK Phosphorylation in Murine and Human Hematopoietic Stem/Progenitor Cells
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DOI：
10.1002/jcp.22507
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发表时间：
2011-07-01
影响因子：
5.6
通讯作者：
Paredes-Gamero, Edgar J.
中科院分区：
文献类型：
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作者：
Leon, Carlos M. M. P.;Barbosa, Christiano M. V.;Paredes-Gamero, Edgar J.
尽管细胞内Ca(2+)(Ca(I)(2+))参与了造血过程，但细胞因子诱导的细胞内途径与Ca(I)(2+)信号之间的关系仍不清楚。本研究探讨了IL-3和GM-CSF刺激的原代小鼠和人造血干/祖细胞中钙离子信号转导通路与细胞外信号调节蛋白1/2(ERK1/2)通路整合的分子机制。我们的结果表明，IL-3和GM-CSF刺激可引起小鼠和人造血干/祖细胞三磷酸肌醇(IP(3))水平和Ca(I)(2+)释放增加。此外，Ca(I)(2+)信号转导抑制剂如肌醇1，4，5-三磷酸受体拮抗剂(2-APB)、PKC抑制剂(GF109203)和CaMKII抑制剂(KN-62)可阻断由IL-3和GM-CSF激活的MEK的磷酸化，提示Ca(2+)依赖的激酶参与了MEK的激活。此外，我们还鉴定了磷脂酶C-γ2(PLC-γ2)是一种磷脂酶C-γ，负责IL-3和GM-CSF诱导造血干/祖细胞释放Ca(2+)。此外，PLCg抑制剂U73122可显著减少细胞因子刺激后粒-巨噬细胞集落形成单位的数量。在小鼠和人的造血干/祖细胞中也得到了类似的结果。综上所述，这些数据表明PLC-Gamma-2和Ca(2+)信号通过调节MEK在小鼠和人类造血干/祖细胞中发挥作用。J.细胞。物理。226：1780-1792,2011。(C)2010年Wiley-Liss公司
Even though the involvement of intracellular Ca(2+) (Ca(i)(2+)) in hematopoiesis has been previously demonstrated, the relationship between Ca(i)(2+) signaling and cytokine-induced intracellular pathways remains poorly understood. Herein, the molecular mechanisms integrating Ca(2+) signaling with the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway in primary murine and human hematopoietic stem/progenitor cells stimulated by IL-3 and GM-CSF were studied. Our results demonstrated that IL-3 and GM-CSF stimulation induced increased inositol 1,4,5-trisphosphate (IP(3)) levels and Ca(i)(2+) release in murine and human hematopoietic stem/ progenitor cells. In addition, Ca(i)(2+) signaling inhibitors, such as inositol 1,4,5-trisphosphate receptor antagonist (2-APB), PKC inhibitor (GF109203), and CaMKII inhibitor (KN-62), blocked phosphorylation of MEK activated by IL-3 and GM-CSF, suggesting the participation of Ca(2+)-dependent kinases in MEK activation. In addition, we identify phospholipase C gamma 2 (PLC gamma 2) as a PLC gamma responsible for the induction of Ca(2+) release by IL-3 and GM-CSF in hematopoietic stem/progenitor cells. Furthermore, the PLCg inhibitor U73122 significantly reduced the numbers of granulocyte-macrophage colony-forming units after cytokine stimulation. Similar results were obtained in both murine and human hematopoietic stem/progenitor cells. Taken together, these data indicate a role for PLC gamma 2 and Ca(2+) signaling through the modulation of MEK in both murine and human hematopoietic stem/ progenitor cells. J. Cell. Physiol. 226: 1780-1792, 2011. (C) 2010 Wiley-Liss, Inc.