Quantitative proteomic analysis of dexamethasone-induced effects on osteoblast differentiation, proliferation, and apoptosis in MC3T3-E1 cells using SILAC.
Quantitative proteomic analysis of dexamethasone-induced effects on osteoblast differentiation, proliferation, and apoptosis in MC3T3-E1 cells using SILAC.
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DOI：
10.1007/s00198-010-1434-8
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发表时间：
2011-07
期刊：
影响因子：
--
通讯作者：
Ge RS
中科院分区：
文献类型：
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作者：
Hong D;Chen HX;Yu HQ;Wang C;Deng HT;Lian QQ;Ge RS
成骨细胞分化障碍是糖皮质激素诱导骨质疏松症（GCOP）的病因之一。利用细胞培养氨基酸稳定同位素标记（SILAC）对地塞米松（DEX）诱导的成骨细胞分化、增殖和凋亡的定量蛋白质组学分析表明，MC3T3-E1细胞中一些关键蛋白发生了剧烈变化。成骨细胞分化障碍是GCOP的主要原因之一。SILAC能够对细胞中蛋白质变化进行精确的定量蛋白质组学分析，以探索GCOP的潜在机制。将骨祖细胞MC3T3-E1分别加入或不加入10−6 M DEX处理7天，观察细胞的分化能力、增殖和凋亡情况。采用液相色谱耦合串联质谱法分析蛋白质水平的变化。在本研究中，10−6 M DEX在第7天抑制成骨细胞的分化和增殖，但诱导成骨细胞MC3T3-E1细胞凋亡。我们发现，10−6 M DEX增加了MC3T3-E1细胞中微管蛋白（TUBA1A、TUBB2B和TUBB5）、IQGAP1、S100蛋白（S100A11、S100A6、S100A4和S100A10）、肌球蛋白蛋白（MYH9和MYH11）、凋亡和应激蛋白的水平，同时抑制了ATP合成酶（atp50、ATP5H、ATP5A1和ATP5F1）、G3BP-1和ras相关蛋白（Rab-1A、Rab-2A和Rab-7）的蛋白水平。ATP合成酶、肌球蛋白蛋白、GTPase小超家族和S100蛋白的一些成员可能参与GCs对成骨祖细胞的功能抑制。这种蛋白表达变化可能在应对GCOP时具有病理意义。
The impairment of osteoblast differentiation is one cause of the glucocorticoid-induced osteoporosis (GCOP). The quantitative proteomic analysis of the dexamethasone (DEX)-induced effects of osteoblast differentiation, proliferation, and apoptosis using stable-isotope labeling by amino acids in cell culture (SILAC) demonstrated drastic changes of some key proteins in MC3T3-E1 cells. The impairment of osteoblast differentiation is one of the main explanations of GCOP. SILAC enables accurate quantitative proteomic analysis of protein changes in cells to explore the underlying mechanism of GCOP. Osteoprogenitor MC3T3-E1 cells were treated with or without 10−6 M DEX for 7 days, and the differentiation ability, proliferation, and apoptosis of the cells were measured. The protein level changes were analyzed using SILAC and liquid chromatography-coupled tandem mass spectrometry. In this study, 10−6 M DEX inhibited both osteoblast differentiation and proliferation but induced apoptosis in osteoprogenitor MC3T3-E1 cells on day 7. We found that 10−6 M DEX increased the levels of tubulins (TUBA1A, TUBB2B, and TUBB5), IQGAP1, S100 proteins (S100A11, S100A6, S100A4, and S100A10), myosin proteins (MYH9 and MYH11), and apoptosis and stress proteins, while inhibited the protein levels of ATP synthases (ATP5O, ATP5H, ATP5A1, and ATP5F1), G3BP-1, and Ras-related proteins (Rab-1A, Rab-2A, and Rab-7) in MC3T3-E1 cells. Several members of the ATP synthases, myosin proteins, small GTPase superfamily, and S100 proteins may participate in functional inhibition of osteoblast progenitor cells by GCs. Such protein expression changes may be of pathological significance in coping with GCOP.