Proteomic profiling of bone marrow mesenchymal stem cells upon transforming growth factor β1 stimulation
Proteomic profiling of bone marrow mesenchymal stem cells upon transforming growth factor β1 stimulation
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DOI：
10.1074/jbc.m407368200
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发表时间：
2004-10-15
影响因子：
4.8
通讯作者：
Li, S
中科院分区：
文献类型：
--
作者：
Wang, DJ;Park, JS;Li, S
骨髓间充质干细胞（MSC）可以分化成不同类型的细胞，在细胞治疗和组织工程方面具有巨大的潜力。转化生长因子β1（TGF-β）在细胞分化和血管重塑中发挥重要作用。我们发现 TGF-β 诱导 MSC 中细胞形态的变化和肌动蛋白纤维的增加。为了确定 TGF-β 对 MSC 的整体影响，我们采用蛋白质组策略来分析 TGF-β 对人 MSC 蛋白质组的影响。通过使用二维凝胶电泳和电喷雾电离与四极杆/飞行时间串联质谱仪联用，我们生成了 MSC 的蛋白质组参考图谱，并鉴定了 30 种类似的蛋白质，它们响应 TGF-β 而表达或磷酸化增加或减少。 TGF-β调节的蛋白质包括细胞骨架蛋白、基质合成蛋白、膜蛋白、代谢酶等。TGF-β增加平滑肌α-肌动蛋白的表达，减少凝溶胶蛋白的表达。凝溶胶蛋白的过表达抑制了 TGF-β 诱导的平滑肌 α-肌动蛋白的组装；另一方面，敲低凝溶胶蛋白的表达增强了α-肌动蛋白和肌动蛋白丝的组装，而不会显着影响α-肌动蛋白的表达。这些结果表明TGF-β协调α-肌动蛋白的增加和凝溶胶蛋白的减少以促进MSC分化。这项研究表明蛋白质组学工具对于研究干细胞分化和阐明潜在的分子机制很有价值。
Bone marrow mesenchymal stem cells (MSCs) can differentiate into different types of cells and have tremendous potential for cell therapy and tissue engineering. Transforming growth factor beta1 (TGF-beta) plays an important role in cell differentiation and vascular remodeling. We showed that TGF-beta induced cell morphology change and an increase in actin fibers in MSCs. To determine the global effects of TGF-beta on MSCs, we employed a proteomic strategy to analyze the effect of TGF-beta on the human MSC proteome. By using two-dimensional gel electrophoresis and electrospray ionization coupled to quadrupole/time-of-flight tandem mass spectrometers, we have generated a proteome reference map of MSCs, and we identified similar to30 proteins with an increase or decrease in expression or phosphorylation in response to TGF-beta. The proteins regulated by TGF-beta included cytoskeletal proteins, matrix synthesis proteins, membrane proteins, metabolic enzymes, etc. TGF-beta increased the expression of smooth muscle alpha-actin and decreased the expression of gelsolin. Overexpression of gelsolin inhibited TGF-beta-induced assembly of smooth muscle alpha-actin; on the other hand, knocking down gelsolin expression enhanced the assembly of alpha-actin and actin filaments without significantly affecting alpha-actin expression. These results suggest that TGF-beta coordinates the increase of alpha-actin and the decrease of gelsolin to promote MSC differentiation. This study demonstrates that proteomic tools are valuable in studying stem cell differentiation and elucidating the underlying molecular mechanisms.