Comparative proteomics of human embryonic stem cells and embryonal carcinoma cells.
Comparative proteomics of human embryonic stem cells and embryonal carcinoma cells.
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DOI：
10.1002/pmic.200900483
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发表时间：
2010-04
期刊：
影响因子：
3.4
通讯作者：
Pandey, Akhilesh
中科院分区：
文献类型：
--
作者：
Chaerkady, Raghothama;Kerr, Candace L.;Kandasamy, Kumaran;Marimuthu, Arivusudar;Gearhart, John D.;Pandey, Akhilesh
多能人类胚胎干细胞（ESC）可以在体外分化成多种有望用于移植治疗的细胞。人类胚胎癌细胞 (ECC) 是人类畸胎瘤的干细胞，被认为是人类 ESC 的近似但恶性的对应物。本研究利用 iTRAQ 方法对 ESC 和 ECC 进行了全面的定量蛋白质组分析。使用二维液相色谱和串联质谱分析，我们鉴定并定量了约 1,800 种蛋白质。其中包括与多能性和发育以及紧密连接信号传导和 TGF β 受体途径相关的蛋白质。 ESC 和 ECC 之间近 200 种蛋白质的丰度差异超过 2 倍。 ESC 中含量较高的早期发育标志物的例子包括 β-半乳糖苷结合凝集素 (LGALS1)、未分化胚胎细胞转录因子-1 (UTF1)、DNA 胞嘧啶甲基转移酶 3 异构体-B (DNMT3B)、黑色素瘤抗原家族-A4 (MAGEA4) 和干扰素诱导跨膜蛋白-1 (IFITM1)。相比之下，CD99 抗原 (CD99)、生长分化因子 3 (GDF3)、细胞视黄酸结合蛋白 2 (CRABP2) 和发育多能性相关蛋白 4 (DPPA4) 是 ECC 中高表达的蛋白。 ECC 中高表达的几种蛋白质，如热休克 27 kDa 蛋白-1 (HSPB1)、丝裂原激活蛋白激酶激酶-1 (MAP3K1)、B 细胞抑制剂样核因子 kappa 轻链多肽基因增强子-2 (NFKBIL2) 和 S100 钙结合蛋白-A4 (S100A4) 也被归因于 其他系统的恶性肿瘤。重要的是，免疫细胞化学用于验证蛋白质子集的蛋白质组分析。总之，这是第一个针对人类 ESC 和 ECC 的大规模定量蛋白质组学研究，它提供了有关这两种密切相关但发育不同的干细胞的调节因子的关键信息。
Pluripotent human embryonic stem cells (ESCs) can be differentiated in vitro into a variety of cells which hold promise for transplantation therapy. Human embryonal carcinoma cells (ECCs), stem cells of human teratocarcinomas, are considered a close but malignant counterpart to human ESCs. In this study, a comprehensive quantitative proteomic analysis of ESCs and ECCs was carried out using the iTRAQ method. Using two-dimensional liquid chromatography and tandem mass spectrometry analyses, we identified and quantitated ~1,800 proteins. Among these are proteins associated with pluripotency and development as well as tight junction signaling and TGF beta receptor pathway. Nearly ~200 proteins exhibit >2 fold difference in abundance between ESCs and ECCs. Examples of early developmental markers high in ESCs include beta-galactoside-binding lectin (LGALS1), undifferentiated embryonic cell transcription factor-1 (UTF1), DNA cytosine methyltransferase 3 isoform-B (DNMT3B), melanoma antigen family-A4 (MAGEA4), and interferon induced transmembrane protein-1 (IFITM1). In contrast, CD99-antigen (CD99), growth differentiation factor-3 (GDF3), cellular retinoic acid binding protein-2 (CRABP2), and developmental pluripotency associated-4 (DPPA4) were among the highly expressed proteins in ECCs. Several proteins that were highly expressed in ECCs such as heat shock 27 kDa protein-1 (HSPB1), mitogen-activated protein kinase kinase-1 (MAP3K1), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor like-2 (NFKBIL2), and S100 calcium-binding protein-A4 (S100A4) have also been attributed to malignancy in other systems. Importantly, immunocytochemistry was used to validate the proteomic analyses for a subset of the proteins. In summary, this is the first large scale quantitative proteomic study of human ESCs and ECCs, which provides critical information about the regulators of these two closely related, but developmentally-distinct, stem cells.
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