The State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China.
J Cell Biochem. 2012 Jun;113(6):1811-9. doi: 10.1002/jcb.24091.
The multipotent mouse F9 embryonic carcinoma cell is an ideal model system to investigate the mechanism of retinoic acid (RA) in cell differentiation and cell growth control and the biochemical basis of early embryonic development. We reported here a proteomics approach to study protein expression changes during the differentiation of F9 cells into the visceral endoderm. F9 cells were incubated with or without RA at 0, 24, 48, and 72 h. Total proteins extracted were separated by two-dimensional electrophoresis (2-DE) and the protein patterns on the gels were comparatively analyzed by computer. Approximately 1,100 protein spots were detected in the F9 proteome, within the pH 3-10 range. Fourteen protein spots which the levels of expression were found to be altered dramatically during the F9 cells differentiating, and were identified by MALDI-TOF MS or ESI-MS/MS. These proteins included metabolism enzymes, HSP60s, RAN, hnRNP K, FUBP1, VDAC1, STI1, and prohibitin. These proteins are involved in cellar metabolism, gene expression regulation, stress response, and apoptosis, respectively. The data from proteomic analyze are consistent with the result obtained from Western blot analysis. This study increases our understanding of the proteomics changes during F9 cells differentiation induced by RA.
多能性小鼠 F9 胚胎癌细胞系是研究维甲酸（RA）在细胞分化和细胞生长调控中的作用机制以及早期胚胎发育的生化基础的理想模型系统。我们在此报告了一种蛋白质组学方法，用于研究 F9 细胞向内脏内胚层分化过程中的蛋白质表达变化。将 F9 细胞在有或没有 RA 的情况下分别孵育 0、24、48 和 72 小时。提取的总蛋白通过二维电泳（2-DE）分离，并通过计算机比较凝胶上的蛋白质图谱进行分析。在 F9 蛋白质组中，在 pH 3-10 范围内检测到约 1100 个蛋白质斑点。在 F9 细胞分化过程中，发现有 14 个蛋白质斑点的表达水平发生了显著变化，并通过 MALDI-TOF MS 或 ESI-MS/MS 进行了鉴定。这些蛋白质包括代谢酶、HSP60s、RAN、hnRNP K、FUBP1、VDAC1、STI1 和抑制素。这些蛋白质分别参与细胞代谢、基因表达调控、应激反应和细胞凋亡。蛋白质组学分析的数据与 Western blot 分析的结果一致。这项研究增加了我们对 RA 诱导的 F9 细胞分化过程中蛋白质组变化的理解。
