解剖学报

解剖学报 ›› 2014, Vol. 45 ›› Issue (4): 507-515.doi: 10.3969/j.issn.0529-1356.2014.04.012

• 肿瘤生物学 • 上一篇    下一篇

二甲基亚砜诱导小鼠红白血病细胞分化的差异蛋白质组学

杨祖立 朱晓芳 施鸣铭 胡繁 赵辅昆 张世馥*   

  1. 浙江理工大学生命科学学院， 杭州 310018
  • 收稿日期:2013-12-11 修回日期:2014-03-14 出版日期:2014-08-06 发布日期:2014-08-06
  • 通讯作者: 张世馥 E-mail:cklzhang@163.com

Comparative proteomic analyses on the differentiation of dimethyl sulfoxide induced murine erythroleukemia cells

YANG Zu-li  ZHU Xiao-fang  SHI Ming-ming  HU Fan  ZHAO Fu-kun  ZHANG Shi-fu*   

  1. Laboratory of College of Life Sciences, Zhejiang SciTech University, Hangzhou 310018, China
  • Received:2013-12-11 Revised:2014-03-14 Online:2014-08-06 Published:2014-08-06
  • Contact: ZHANG Shi-fu E-mail:cklzhang@163.com

摘要：

目的 利用二甲基亚砜（DMSO）诱导小鼠红白血病（MEL）细胞分化，系统地分析和鉴定诱导分化不同时间的差异蛋白质组学。方法 DMSO诱导MEL细胞分化，通过联苯胺染色、MTT比色法和Ter119免疫荧光等实验检测细胞分化的比率和细胞的活力，并利用双向电泳耦联质谱技术，结合生物信息学分析诱导MEL细胞分化过程中差异表达的蛋白质。结果 1.2% DMSO诱导MEL细胞分化，分别培养0h、6h、12h、24h、36h、48h、72h、96h、120h，利用双向电泳和质谱分析，共鉴定出87种蛋白质。1.2% DMSO作用MEL细胞120h后，细胞诱导分化率达到67%。 MTT实验显示，1.0%、1.2%、1.4% 的DMSO对MEL细胞的活力无明显的抑制作用。结论 DMSO对MEL细胞有诱导分化作用，但无明显地抑制细胞增殖。87种双向电泳差异蛋白质按功能可分为12类，比例最大的前3类分别是：占41% 的酶类蛋白、15% 的结构蛋白、13% 的调节蛋白。

关键词: 小鼠红白血病细胞, 二甲基亚砜, 诱导, 分化, 双向电泳, 小鼠

Abstract:

Objective To explore the differentially expressed proteins during erythroid differentiation. Methods The murine erythroleukemia (MEL) cell were treated by DMSO, and the comparative proteomic was systematically analyzed and identified on different differentiating time points. ratio of cell differentiation and viability were detected by benzidine staining, MTT assay and Ter119 immunofluorescence. Using two-dimensional gel electrophoresis combined with mass spectrometry technology and bioinformatics analysis, we conducted a comparative proteomic analysis on MEL cells during the process of induced differentiation to screen and identify differential proteins. Results The MEL cells induced by 1.2% DMSO for 0 hour, 6hours, 12hours, 24hours, 36hours, 48hours, 72 hours, 96 hours, 120 hours were collected for proteomic analysis, by two-dimensional gel electrophoresis combined with mass spectrometry. A total of 87 kinds of proteins were successfully identified. MEL cells exposed to DMSO at a final concentration of 1.2% for 120 hours reached the highest differentiation rate of 67%. MTT assay showed that 1.0%, 1.2%, 1.4% DMSO had no inhibiting effect on cell vitality. Conclusion DMSO may induce MEL cells to differentiate and have no inhibiting effect on cell vitality. The 87 kinds of differentially expressed proteins from two-dimentional gel electrophoresis may be divided into twelve categories; the most three parts are 41% enzyme protein, 15% structural protein and 13% regulatory protein.

Key words: Murine erythroleukemia cell, Dimethyl sulfoxide, Induce, Differentiation, Two-dimentional gel electrophoresis, Mouse