Heat Shock Protein 90 Stimulates Rat Mesenchymal Stem Cell Migration via PI3K/Akt and ERK1/2 Pathways
Heat Shock Protein 90 Stimulates Rat Mesenchymal Stem Cell Migration via PI3K/Akt and ERK1/2 Pathways
复制标题
DOI：
10.1007/s12013-014-0228-6
复制
发表时间：
2015-01-01
影响因子：
2.6
通讯作者：
Wang, Jianan
中科院分区：
文献类型：
--
作者：
Gao, Feng;Hu, Xinyang;Wang, Jianan
本研究的目的是确定Hsp90 α在调节间充质干细胞（MSCs）迁移中的作用，并探讨这种作用的潜在机制。通过伤口愈合实验和跨井迁移实验评估MSCs的迁移。通过siRNA （sirHsp90 α）抑制MSC中Hsp90 α的表达。采用明胶酶谱法、Western blot分析和实时荧光定量PCR检测骨髓间质干细胞中分泌金属蛋白酶MMP-2和MMP-9的活性及表达水平。实时荧光定量PCR检测VCAM-1和CXCR4细胞因子的基因表达。Akt和ERK的活性通过Western blotting分析，抗体针对这些蛋白的磷酸化形式。用Hsp90 α处理可显著增强MSC迁移，而用sirHsp90 α转染MSC可阻断这种作用。用重组人Hsp90 α (rhHsp90 α)处理细胞后，MMP-2和MMP-9的基因表达和蛋白水平以及它们的分泌和活性均有所提高。与rhHsp90 α孵育的MSC显示CXCR4和VCAM-1基因表达增加。最后，rhHsp90 α处理后，磷酸化Akt和Erk的水平显著升高。这些发现表明，Hsp90 α通过PI3K/Akt和ERK信号通路促进MSCs迁移，这种作用可能是由MMPs、SDF-1/CXCR4通路和VCAM-1介导的。
The objective of this study was to determine the role of Hsp90 alpha in regulating the migration of mesenchymal stem cells (MSCs) and to investigate the underlying mechanisms of this effect. MSCs migration was assessed by wound healing assay and transwell migration assay. Hsp90 alpha expression was silenced in MSC by siRNA (sirHsp90 alpha). The activity of secreted metalloproteases MMP-2 and MMP-9, and their expression levels in MSC were evaluated using gelatin zymography, Western blot analysis and real-time PCR. Gene expression of VCAM-1 and CXCR4 cytokines was evaluated by real-time PCR. Akt and ERK activity were analyzed by Western blotting using antibodies against phosphorylated forms of these proteins. Treatment with Hsp90 alpha significantly enhanced MSC migration, and this effect was blocked by transfecting MSC with sirHsp90 alpha. Treating the cells with recombinant human Hsp90 alpha (rhHsp90 alpha) enhanced gene expression and protein levels of MMP-2 and MMP-9, as well as their secretion and activity. MSC incubated with rhHsp90 alpha exhibited increased gene expression of CXCR4 and VCAM-1. Finally, the levels of phosphorylated Akt and Erk were markedly increased by rhHsp90 alpha treatment. These findings indicate that Hsp90 alpha promotes MSCs migration via PI3K/Akt and ERK signaling pathways, and that this effect is possibly mediated by MMPs, SDF-1/CXCR4 pathway, and VCAM-1.