Division of Molecular and Life Science, Hanyang University, Ansan, Republic of Korea.
Toxicol In Vitro. 2010 Sep;24(6):1696-704. doi: 10.1016/j.tiv.2010.05.017. Epub 2010 May 26.
This study used human embryonic carcinoma (NCCIT) cells to evaluate genotoxicity and other effects of ethanol in earlier stages of cellular development. This undifferentiated pluripotent cell line has unlimited self-renewal capacity and has been shown to differentiate in vitro. We analyzed proteome expression profile of ethanol-treated NCCIT cells and NCCIT cell-derived embryoid bodies (EBs) by MALDI-TOF MS. To test the role of ethanol as an embryotoxic and/or teratogenic factor, MetaCore pathway analysis software (GeneGO) was used to evaluate the process of normal growth and differentiation of NCCIT cells and EBs. We compared the different protein expression profiles of ethanol-treated versus untreated NCCIT cells and NCCIT cell-derived EBs. The ethanol-treated NCCIT cells demonstrate significant up regulation of SMAP1, dual specificity phosphatase 1 and pro isomerase domain-containing 1, cytokeratin 18, triosephosphate isomerase and beta-tubulin. However, ethanol-treated NCCIT cell-derived EBs exhibited upregulated signatures of different proteins, including CDC25B phosphatase, alpha-enolase, 3-phosphoglycerate dehydrogenase and tumor suppressor patched L' isoform, which suggests that ethanol may play a different role in EBs. These proteins exert their function on transcriptional and translational processes. Moreover, the functional proteomic analysis confirms the relationship between ethanol and ethanol-regulated genes and various signaling pathways and networks. The data presented in this study contribute toward the understanding of the molecular mechanisms of ethanol in NCCIT cells and NCCIT cell-derived EBs.
本研究使用人胚胎癌细胞(NCCIT)来评估乙醇在细胞发育早期的遗传毒性和其他作用。这种未分化的多能细胞系具有无限的自我更新能力,并已被证明可以在体外分化。我们通过 MALDI-TOF MS 分析了乙醇处理的 NCCIT 细胞和 NCCIT 细胞衍生的类胚体(EBs)的蛋白质组表达谱。为了测试乙醇作为胚胎毒性和/或致畸性因素的作用,使用 MetaCore 途径分析软件(GeneGO)来评估 NCCIT 细胞和 EBs 的正常生长和分化过程。我们比较了乙醇处理的 NCCIT 细胞与未处理的 NCCIT 细胞和 NCCIT 细胞衍生的 EBs 的不同蛋白质表达谱。乙醇处理的 NCCIT 细胞表现出 SMAP1、双特异性磷酸酶 1 和前异构酶结构域包含蛋白 1、细胞角蛋白 18、磷酸丙糖异构酶和微管蛋白β的显著上调。然而,乙醇处理的 NCCIT 细胞衍生的 EBs 表现出不同蛋白质的上调特征,包括 CDC25B 磷酸酶、α-烯醇酶、3-磷酸甘油酸脱氢酶和肿瘤抑制因子 patched L' 异构体,这表明乙醇在 EBs 中可能发挥不同的作用。这些蛋白质在转录和翻译过程中发挥作用。此外,功能蛋白质组学分析证实了乙醇与乙醇调节基因以及各种信号通路和网络之间的关系。本研究中提供的数据有助于理解乙醇在 NCCIT 细胞和 NCCIT 细胞衍生的 EBs 中的分子机制。