SH2B1β (SH2-Bβ) enhances expression of a subset of nerve growth factor-regulated genes important for neuronal differentiation including genes encoding urokinase plasminogen activator receptor and matrix metalloproteinase 3/10
SH2B1β (SH2-Bβ) enhances expression of a subset of nerve growth factor-regulated genes important for neuronal differentiation including genes encoding urokinase plasminogen activator receptor and matrix metalloproteinase 3/10
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DOI：
10.1210/me.2007-0384
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发表时间：
2008-02-01
影响因子：
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通讯作者：
Carter-Su, Christin
中科院分区：
文献类型：
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作者：
Chen, Linyi;Maures, Travis J.;Carter-Su, Christin
先前的研究表明，适配蛋白SH2B (SH2B1) （SH2-B）与神经生长因子（NGF）受体TrkA的活化形式结合，对NGF依赖性神经突的生长和维持至关重要。为了鉴定对神经突生长至关重要的SH2B β调控基因，我们对对照PC12细胞和稳定过表达SH2B β (PC12-SH2B β)或显性负表达SH2B β (R555E)的PC12细胞进行了微阵列分析。ngf诱导的Plaur和Mmp10基因的微阵列表达在PC12-SH2B β细胞中显著增强，而在PC12-SH2B β (R555E)细胞中，ngf诱导的Plaur和Mmp3基因的微阵列表达则显著降低。Plaur、Mmp3和Mmp10是在NGF作用6小时后表达上调最多的12个基因之一。它们的蛋白产物[尿激酶纤溶酶原激活物受体（uPAR），基质金属蛋白酶3 （MMP3）和MMP10]处于相同的细胞外基质降解途径；uPAR先前已被证明对ngf诱导的神经突生长至关重要。实时荧光定量PCR分析显示，在PC12-SH2B β (R555E)细胞中，使用短发夹RNA抑制SH2Band抑制内源性sh2b时，SH2B β增强了这三个基因对NGF的诱导，抑制了这三个基因对NGF的诱导。在PC12-SH2B β细胞中，ngf诱导的uPAR和MMP3/10水平以及通过Matrigel （mmp3依赖）的神经突生长也增加。这些结果表明，SH2B β刺激ngf诱导的神经元分化至少部分是通过增强神经突起生长所需的特定ngf敏感基因亚群的表达，包括Plaur、Mmp3和/或Mmp10。
Previous work showed that the adapter protein SH2B adapter protein beta (SH2B1) (SH2-B) binds to the activated form of the nerve growth factor (NGF) receptor TrkA and is critical for both NGF-dependent neurite outgrowth and maintenance. To identify SH2B beta-regulated genes critical for neurite outgrowth, we performed microarray analysis of control PC12 cells and PC12 cells stably overexpressing SH2B beta (PC12-SH2B beta) or the dominant-negative SH2B beta(R555E) [PC12-SH2B beta(R555E)]. NGF-induced microarray expression of Plaur and Mmp10 genes was greatly enhanced in PC12-SH2B beta cells, whereas NGF-induced Plaur and Mmp3 expression was substantially depressed in PC12-SH2B beta(R555E) cells. Plaur, Mmp3, and Mmp10 are among the 12 genes most highly upregulated after 6 h of NGF. Their protein products [urokinase plasminogen activator receptor (uPAR), matrix metalloproteinase 3 (MMP3), and MMP10] lie in the same pathway of extracellular matrix degradation; uPAR has been shown previously to be critical for NGF-induced neurite outgrowth. Quantitative real-time PCR analysis revealed SH2B beta enhancement of NGF induction of all three genes and the suppression of NGF induction of all three when endogenous SH2Bwas reduced using short hairpin RNA against SH2Band in PC12-SH2B beta(R555E) cells. NGF-induced levels of uPAR and MMP3/10 and neurite outgrowth through Matrigel (MMP3-dependent) were also increased in PC12-SH2B beta cells. These results suggest that SH2B beta stimulates NGF-induced neuronal differentiation at least in part by enhancing expression of a specific subset of NGF-sensitive genes, including Plaur, Mmp3, and/or Mmp10, required for neurite outgrowth.