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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Proteome alteration of early-stage differentiation of mouse embryonic stem cells into hepatocyte-like cells.
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Proteome alteration of early-stage differentiation of mouse embryonic stem cells into hepatocyte-like cells.

机译：小鼠胚胎干细胞早期分化为肝细胞样细胞的蛋白质组学改变。

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摘要

To explore the molecular basis of inducible differentiation of embryonic stem cells into hepatocyte-like cells, a proteomic strategy was utilized to examine the global protein expression alterations after early-stage differentiation of a mouse D3 embryonic stem (ES) cell line along hepatic lineage. The undifferentiated D3 cells were treated stepwise with combinations of defined chemicals and growth factors. The differentiated cells were identified by hepatocyte-like morphology, expressed liver-specific markers as well as the evidence of glycogen storage. The subsequent proteomic separation and identification were performed with 2-DE followed by MALDI-TOF-MS/MS analysis. Of the 119 differentially displayed protein spots analyzed, 90 spots presenting 64 distinct proteins were finally identified. The interested protein expressions were validated by Western blotting such as albumin and cytokeratin-8. Bioinformatic annotations indicated that this set of proteins was enriched with transcription, translation regulation and protein processing, energy/metabolism and chaperone functions. A part of them had been found to be involved in the differentiation of mouse ES cells. Interestingly, approximately 40% of these proteins had been previously reported as being dysregulated in hepatocellular carcinoma. It suggested that these changed proteins may be candidate regulators of ES cell differentiation, some of them may be specific to hepatic differentiation.
机译：为了探索可诱导分化的胚胎干细胞分化为肝细胞样细胞的分子基础，蛋白质组学策略被用来检查小鼠D3胚胎干（ES）细胞沿肝谱系早期分化后的整体蛋白表达变化。将未分化的D3细胞逐步用确定的化学物质和生长因子组合处理。通过肝细胞样形态鉴定分化的细胞，表达肝特异性标记以及糖原储存的证据。随后用2-DE进行蛋白质组分离和鉴定，然后进行MALDI-TOF-MS / MS分析。在分析的119个差异显示的蛋白质斑点中，最终鉴定出了代表64种不同蛋白质的90个斑点。通过蛋白质印迹，例如白蛋白和细胞角蛋白8，验证了感兴趣的蛋白质表达。生物信息学注释表明，这组蛋白质富含转录，翻译调控和蛋白质加工，能量/代谢和伴侣功能。已经发现它们的一部分与小鼠ES细胞的分化有关。有趣的是，先前已报道这些蛋白质中约40％在肝细胞癌中失调。这表明这些改变的蛋白质可能是ES细胞分化的候选调节剂，其中一些可能对肝细胞分化具有特异性。

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