Involvement of Cytoskeleton-associated Proteins in the Commitment of C3H10T1/2 Pluripotent Stem Cells to Adipocyte Lineage Induced by BMP2/4
Involvement of Cytoskeleton-associated Proteins in the Commitment of C3H10T1/2 Pluripotent Stem Cells to Adipocyte Lineage Induced by BMP2/4
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细胞骨架相关蛋白参与 BMP2/4 诱导的 C3H10T1/2 多能干细胞向脂肪细胞谱系的定向
DOI：
10.1074/mcp.m110.002691
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发表时间：
2011-01-01
影响因子：
7
通讯作者：
Tang, Qi-Qun
中科院分区：
文献类型：
--
作者：
Huang, Hai-Yan;Hu, Ling-Ling;Tang, Qi-Qun
产生成熟脂肪细胞的发育途径涉及两个不同的阶段：定型和终末分化。虽然有助于终末脂肪细胞分化的重要蛋白质/因子已经被很好地定义，但是在间充质干细胞向脂肪细胞谱系细胞的定型中的蛋白质/因子还没有。在这项研究中，我们应用蛋白质组学分析来表征未定向的C3 H10 T1/2多能干细胞与那些已通过BMP 4或BMP 2定向为脂肪细胞谱系的细胞之间的差异，目的是识别此类蛋白质/因子并了解分子机制控制脂肪细胞谱系定向的最早阶段。发现8种蛋白质被BMP 2上调，27种蛋白质被BMP 4上调，而5种独特的蛋白质被BMP 2/4上调至少10倍，包括3种细胞凋亡相关蛋白质（即赖氨酰氧化酶（LOX）、转录控制肿瘤蛋白1（TPT 1）和α B-晶状体蛋白）。Western blotting进一步证实了BMP 2/4诱导的C3 H10 T1/2中这些细胞因子相关蛋白的表达。重要的是，LOX表达的敲低完全阻止了定型，而TPT 1和α B-晶体蛋白表达的敲低部分抑制了定型。一些已发表的报告表明，细胞形状可以影响部分定向的脂肪细胞，成骨细胞和软骨细胞的前体细胞的分化。我们观察到一个戏剧性的变化，细胞形状的承诺过程中，我们表明，敲低这些cytokereon相关蛋白阻止细胞形状的变化，并恢复F-肌动蛋白组织成应力纤维和抑制承诺的脂肪细胞谱系。我们的研究表明，这些差异表达的细胞因子相关蛋白可能决定了间充质干细胞的命运，通过细胞形状调节致力于脂肪细胞谱系。Molecular & Cellular Proteomics 10：10.1074/mcp. M110.002691，1-8，2011.
The developmental pathway that gives rise to mature adipocytes involves two distinct stages: commitment and terminal differentiation. Although the important proteins/factors contributing to terminal adipocyte differentiation have been well defined, the proteins/factors in the commitment of mesenchymal stem cells to the adipocyte lineage cells have not. In this study, we applied proteomics analysis profiling to characterize differences between uncommitted C3H10T1/2 pluripotent stem cells and those that have been committed to the adipocyte lineage by BMP4 or BMP2 with the goal to identify such proteins/factors and to understand the molecular mechanisms that govern the earliest stages of adipocyte lineage commitment. Eight proteins were found to be up-regulated by BMP2, and 27 proteins were up-regulated by BMP4, whereas five unique proteins were up-regulated at least 10-fold by both BMP2/4, including three cytoskeleton-associated proteins (i.e. lysyl oxidase (LOX), translationally controlled tumor protein 1 (TPT1), and alpha B-crystallin). Western blotting further confirmed the induction of the expression of these cytoskeleton-associated proteins in the committed C3H10T1/2 induced by BMP2/4. Importantly, knockdown of LOX expression totally prevented the commitment, whereas knockdown of TPT1 and alpha B-crystallin expression partially inhibited the commitment. Several published reports suggest that cell shape can influence the differentiation of partially committed precursors of adipocytes, osteoblasts, and chondrocytes. We observed a dramatic change of cell shape during the commitment process, and we showed that knockdown of these cytoskeleton-associated proteins prevented the cell shape change and restored F-actin organization into stress fibers and inhibited the commitment to the adipocyte lineage. Our studies indicate that these differentially expressed cytoskeleton-associate proteins might determine the fate of mesenchymal stem cells to commit to the adipocyte lineage through cell shape regulation. Molecular & Cellular Proteomics 10: 10.1074/mcp.M110.002691, 1-8, 2011.