Laboratory of Molecular Signaling and Stem Cells Therapy, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology, Beijing, China.
Department of Implant Dentistry, Capital Medical University School of Stomatology, Beijing, China.
J Cell Biochem. 2018 Nov;119(10):8450-8459. doi: 10.1002/jcb.27069. Epub 2018 Jul 16.
To investigate the function of epiregulin (EREG) in the migration and chemotaxis ability of mesenchymal stem cells. Adipose-derived stem cells (ADSCs) were used in this investigation. Lentiviral EREG short hairpin RNA was applied to silence EREG expression in ADSCs. Human recombinant EREG protein (rhEREG) was used to perform a gain-of-function study. Scratch-simulated wound migration and transwell chemotaxis assays were used to examine the migration and chemotaxis capacity of ADSCs in vitro. Using a Western blot assay, the expressions of p38 mitogen-activated protein kinase (p38 MAPK), c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinases 1 and 2 (Erk1/2), and protein kinase B were detected. Depletion of EREG caused by specific short hairpin RNA restrained the migration and chemotaxis ability of ADSCs and inhibited the expressions of phosphorylated p38 MAPK, JNK, and Erk1/2. rhEREG improved ADSCs migration and chemotaxis capacity, which was repressed by knockdown of EREG and rescued the expressions of phosphorylated p38 MAPK, JNK, and Erk1/2 impaired by silencing EREG. Furthermore, rhEREG-improved migration and chemotaxis ability in EREG-depleted-ADSCs was restricted by a specific inhibitor, SB203580, for blocking p38 MAPK signaling, PD98059 for blocking Erk1/2 signaling, or SP600125 for blocking JNK signaling in ADSCs separately. EREG promotes migration and chemotaxis ability of ADSCs through MAPK signaling pathways.
探讨表皮调节素（EREG）在间充质干细胞（MSCs）迁移和趋化能力中的作用。本研究采用脂肪来源的干细胞（ADSCs）。应用慢病毒 EREG 短发夹 RNA 沉默 ADSCs 中的 EREG 表达。采用人重组 EREG 蛋白（rhEREG）进行功能获得研究。划痕模拟的体外迁移和 Transwell 趋化实验检测 ADSCs 的体外迁移和趋化能力。采用 Western blot 检测 p38 丝裂原活化蛋白激酶（p38 MAPK）、c-Jun N 末端激酶（JNK）、细胞外信号调节激酶 1 和 2（Erk1/2）和蛋白激酶 B 的表达。特异性短发夹 RNA 沉默 EREG 导致 ADSCs 的迁移和趋化能力受到抑制，并抑制磷酸化 p38 MAPK、JNK 和 Erk1/2 的表达。rhEREG 改善 ADSCs 的迁移和趋化能力，而 EREG 沉默则抑制了 EREG 表达，并挽救了 EREG 沉默所损害的磷酸化 p38 MAPK、JNK 和 Erk1/2 的表达。此外，rhEREG 在 EREG 耗竭的 ADSCs 中改善迁移和趋化能力的作用受到特异性抑制剂的限制，包括阻断 p38 MAPK 信号通路的 SB203580、阻断 Erk1/2 信号通路的 PD98059 或阻断 JNK 信号通路的 SP600125。EREG 通过 MAPK 信号通路促进 ADSCs 的迁移和趋化能力。
