Fibroblast growth factor-2 stimulates proliferation of human adipose-derived stem cells via Src activation
Fibroblast growth factor-2 stimulates proliferation of human adipose-derived stem cells via Src activation
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DOI：
10.1186/s13287-019-1462-z
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发表时间：
2019-11-27
影响因子：
7.5
通讯作者：
Kusumoto, Kenji
中科院分区：
文献类型：
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作者：
Ma, Yuanyuan;Kakudo, Natsuko;Kusumoto, Kenji
研究背景人脂肪源性干细胞（human adipose-derived stem cells，hASCs）是间充质干细胞（mesenchymal stem cells，MSCs）的一个亚群，被认为是最有前途的干细胞之一。我们先前发现成纤维细胞生长因子-2（FGF-2）促进hASC的增殖和分化。然而，FGF-2参与hASC生长的机制尚未研究。方法将人脂肪干细胞（hASCs）与成纤维细胞生长因子2（FGF-2）共同培养，观察细胞生长情况。观察FGF受体（FGFR）抑制剂（NVP-BGJ 398）、ERK 1/2抑制剂（PD 98059）、PI 3 K/Akt抑制剂（LY 294002）、JNK抑制剂（SP 600125）、p38 MAPK抑制剂（SB 203580）和Src抑制剂（PP 1）对细胞增殖的影响。同时，我们在蛋白水平上评估了FGFR抑制剂对几种信号传导酶如ERK 1/2、JNK、p38和Akt的影响。还检查了FGF-2对Src活化的参与。结果FGF-2在浓度低于10 ng/ml时可明显促进hASCs的增殖，并促进细胞向S期和G2/M期进展。增殖被FGFR抑制剂（NVP-BGJ 398）和各种信号传导途径抑制剂（例如Erk 1/2抑制剂（PD 98059）、PI 3 K/Akt抑制剂（LY 294002）、JNK抑制剂（SP 600125）和p38 MAPK抑制剂（SB 203580））阻断。FGFR抑制剂降低了几种信号传导途径中蛋白激酶的活化，如AKT、Erk 1/2、JNK和p38。FGFR的下游激酶Src被FGF-2激活，其激活被FGFR抑制剂取消。MEK 1/2是Src的下游激酶，受FGF-2的调控。Src抑制剂（PP 1）通过抑制Src和MEK 1/2显著阻断hASCs的增殖。结论Src的激活是FGF-2介导ASCs增殖以及随后多种信号通路激活的必要条件。
Background Human adipose-derived stem cells (hASCs) are a subset of mesenchymal stem cells (MSCs); it has been regarded as one of the most promising stem cells. We previously found that fibroblast growth factor-2 (FGF-2) enhanced the proliferation and differentiation of hASC. However, the mechanisms involved in the growth of hASCs by FGF-2 have not been investigated. Methods Human adipose-derived stem cells (hASCs) were cultured with FGF-2, and cell growth was assessed. Effects of FGF Receptor (FGFR) inhibitor (NVP-BGJ398), ERK1/2 inhibitor (PD98059), PI3K/Akt inhibitor (LY294002), JNK inhibitor (SP600125), and p38 MAPK inhibitor (SB203580) and Src inhibitor (PP1) on the proliferation were investigated. At the same time, we assessed the effect of FGFR inhibitor on several signaling enzymes such as ERK1/2, JNK, p38, and Akt, in protein level. The involvement of Src activation by FGF-2 was also examined. Results FGF-2 markedly promoted proliferation of hASCs at concentrations lower than 10 ng/ml and stimulated cell progression to the S and G2/M phases. Proliferation was blocked by the FGFR inhibitor (NVP-BGJ398) and various signaling pathway inhibitors, such as Erk1/2 inhibitor (PD98059), PI3K/Akt inhibitor (LY294002), JNK inhibitor (SP600125), and p38MAPK inhibitor (SB203580). The FGFR inhibitor reduced the activation of protein kinases, such as AKT, Erk1/2, JNK, and p38, in several signaling pathways. The downstream kinase of FGFR, Src, was activated by FGF-2, and its activation was canceled by the FGFR inhibitor. MEK1/2, a downstream kinase of Src, was parallelly regulated by FGF-2. The Src inhibitor (PP1) markedly blocked the proliferation of hASCs via inhibition of Src and MEK1/2. Conclusion Src activation is indispensable for FGF-2-mediated proliferation of ASCs, as well as the subsequent activation of multi-signaling pathways.