IGF-I stimulates chemotaxis of human neuroblasts. Involvement of type 1 IGF receptor, IGF binding proteins, phosphatidylinositol-3 kinase pathway and plasmin system.
IGF-I stimulates chemotaxis of human neuroblasts. Involvement of type 1 IGF receptor, IGF binding proteins, phosphatidylinositol-3 kinase pathway and plasmin system.
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DOI：
10.1677/joe.0.1650123
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发表时间：
2000-04
期刊：
影响因子：
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通讯作者：
A. Puglianiello;D. Germani;P. Rossi;S. Cianfarani
中科院分区：
文献类型：
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作者：
A. Puglianiello;D. Germani;P. Rossi;S. Cianfarani
SH-SY5Y人神经母细胞瘤细胞表达IGF受体、IGF和IGF结合蛋白（igfbp），为研究IGF系统在人神经元发育中的作用提供了模型。我们通过细胞迁移实验研究了IGF-I和des(1-3)IGF-I对SH-SY5Y细胞运动的影响，该实验基于对跨越8微米孔径膜和琼脂糖滴周围迁移的细胞数量的评估。IGF-I和des(1-3)IGF-I以剂量依赖的方式刺激神经母细胞趋化。用这些药物处理细胞24小时后，IGF-I显著增加70%，des(1-3)IGF-I显著增加90%；P < 0。0001)相对于控制条件的细胞运动。添加抗1型IGF受体（α - ir3）单克隆抗体，可显著（P<0.05）降低IGF- i诱导的细胞活力（降低30%）和des(1-3)IGF- i诱导的细胞活力（降低70%）。Wortmannin是一种磷脂酰肌醇(PI)-3激酶细胞内信号传导的特异性抑制剂，也能减少IGF刺激的细胞迁移（超过40%，P<0.01），表明PI-3激酶途径在介导IGF对神经母细胞迁移的作用中起关键作用。最后，用纤溶酶原（PLG）处理的细胞显著增强了神经母细胞的迁移（超过200%，P<0.01），而用PLG抑制剂4-（2-氨基乙基）-苯磺酰氟（4-（2-氨基乙基）-苯磺酰氟）处理的细胞降低了细胞的运动（80%，P<0.01），这表明PLG依赖性IGFBP蛋白水解参与了神经母细胞运动的调节。综上所述，IGF- 1通过激活1型IGF受体和PI-3激酶细胞内通路，是神经母细胞迁移的有效刺激物。igfbp和纤溶酶系统似乎在细胞运动中发挥作用，尽管它们参与的性质和程度尚未阐明。
SH-SY5Y human neuroblastoma cells express IGF receptors, IGFs and IGF binding proteins (IGFBPs), and provide a model for studying the role of the IGF system in human neuronal development. We investigated the effect of IGF-I and des(1-3)IGF-I on the motility of SH-SY5Y cells by a cell migration assay based on the assessment of the number of cells which migrated across 8 microm pore size membranes and around an agarose drop. IGF-I and des(1-3)IGF-I stimulated neuroblast chemotaxis in a dose-dependent manner. Treatment of cells with these agents for 24 h resulted in a significant increase (IGF-I by 70% and des(1-3)IGF-I by 90%; P<0. 0001) in cell motility relative to control conditions. Addition of monoclonal antibody against type 1 IGF receptor (alpha-IR3), significantly (P<0.05) reduced the cell motility induced by IGF-I (by 30%) and des(1-3)IGF-I (by 70%). Wortmannin, a specific inhibitor of phosphatidylinositol (PI)-3 kinase intracellular signalling, also reduced the IGF-stimulated cell migration (by over 40%, P<0.01), indicating a key role of the PI-3 kinase pathway in mediating the IGF effect on neuroblast migration. Finally, cell treatment with plasminogen (PLG) markedly enhanced neuroblast migration (by over 200%, P<0.01), whereas incubation with the PLG inhibitor 4-(2-aminoethyl)-benzenesulphonyl fluoride reduced cell motility (by 80%, P<0.01), thus suggesting an involvement of PLG-dependent IGFBP proteolysis in the regulation of neuroblast motility. In conclusion, IGF-I is a potent stimulator of neuroblast migration through the activation of type 1 IGF receptor and the PI-3 kinase intracellular pathway. IGFBPs and the plasmin system seem to play a role in cell motility, although the nature and the extent of their involvement has yet to be elucidated.