解剖学报

解剖学报 ›› 2014, Vol. 45 ›› Issue (5): 670-674.doi: 10.3969/j.issn.0529-1356.2014.05.015

• 细胞和分子生物学 • 上一篇    下一篇

p38-2G4蛋白高表达对小鼠红白血病细胞增殖和分化的影响

朱晓芳 施鸣铭 杨祖立 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:2014-01-02 修回日期:2014-03-14 出版日期:2014-10-06 发布日期:2013-10-06
  • 通讯作者: 张世馥 E-mail:cklzhang@163.com

Effects of p38-2G4 high-expression on the proliferation and erythriod differentiateon of murine erythroleukemia cells

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

  1. College of Life Sciences, Zhejiang SciTech University,Hangzhou 310018, China
  • Received:2014-01-02 Revised:2014-03-14 Online:2014-10-06 Published:2013-10-06
  • Contact: ZHANG Shi-fu E-mail:cklzhang@163.com

摘要：

目的 探讨小鼠增殖相关蛋白p38-2G4基因表达上调对小鼠红白血病（MEL）细胞增殖和诱导分化能力的影响。方法 构建p38-2G4-pLJM1高表达重组载体与pCMV-VSV-G和pCMV-dR8.2包装质粒共转染HEK293T细胞慢病毒，感染MEL细胞，建立稳定高表达p38-2G4蛋白的MEL细胞系，Western blotting检测稳定株p38-2G4蛋白的表达。MTT实验和联苯胺染色法检测p38-2G4蛋白高表达对MEL细胞增殖和诱导分化能力的影响。 结果 高表达稳定株中p38-2G4蛋白表达量明显高于对照组细胞（P<0.05）。p38-2G4蛋白的高表达能显著影响MEL诱导分化过程，导致血红蛋白合成减少（P<0.05），但不能明显改变细胞活力（P>0.05）。结论 p38-2G4蛋白的高表达对MEL细胞增殖无明显影响，但能显著影响其被丁酸钠诱导分化的过程。

关键词: p38-2G4, 小鼠红白血病, 慢病毒转染, 高表达, 免疫印迹法, 小鼠

Abstract:

Objective To explore the effect of mouse proliferation-associated protein 2G4 (p38-2G4) high-expression on the proliferation and erythriod differentiation of murine erythroleukemia (MEL) cells. Methods To establish the recombinant lentivirus vector p38-2G4-pLJM1, the p38-2G4-pLJM1 was cotransfected into HEK293T cells to obtain lentivirus with pCMV-VSV-G and pCMV-dR8.2. Lentivirus were infected into MEL cells to establish the stably p38-2G4 high-expressed MEL cells. Western blotting was used to analyse the high-expression efficiency. MTT assay and benzidine staining were applied to detect the cell viability and hemoglobin synthesis of the stable cell line in presence/absence of inducers. Results Western blotting showed that the p38-2G4 high-expression stable cell strain had a higher expression of p38-2G4 as compared to the control group (MEL)（P<0.05）. MTT result showed that there was no difference between the p38-2G4 high-expression cell strain and the control group（P>0.05）, but the hemoglobin synthesis had been reduced as compared to the control group（P<0.05）. Conclusion p38-2G4 high-expression does not affect the cell viability of MEL cells, but inhibits the erythriod differentiation of MEL cells in three independent experiments.

Key words: p38-2G4, Murine erythroleukemia, Lentiviral transfection, High-expression, Western blotling, Mouse