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山东大学学报(医学版) ›› 2011, Vol. 49 ›› Issue (6): 53-58.

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Wnt/β-catenin信号通路在胚胎癌细胞增殖中作用及机制研究

张帅,李毅,武玉玲,史琨,邴鲁军,郝晶   

  1. 山东大学医学院组胚教研室, 济南 250012
  • 收稿日期:2010-11-09 出版日期:2011-06-10 发布日期:2011-06-10
  • 通讯作者: 郝晶(1971- ),女,教授,硕士生导师,主要从事干细胞生物学和生殖生物学研究。 E-mail:haojing@sdu.edu.cn
  • 作者简介:张帅(1984- ),女,硕士研究生,主要从事生殖肿瘤的基础研究。
  • 基金资助:

    国家自然科学基金资助项目(30871405,30971653)。

Effect and mechanism of the Wnt/β-catenin pathway on  proliferation of embryonic carcinoma cells

ZHANG Shuai, LI Yi, WU Yu-ling, SHI Kun, BING Lu-jun, HAO Jing   

  1. Department of Histology and Embryology, School of Medicine, Shandong University, Jinan 250012, China
  • Received:2010-11-09 Online:2011-06-10 Published:2011-06-10

摘要:

目的     探讨Wnt/β-catenin信号通路在胚胎癌细胞增殖中的作用及其机制。方法     取小鼠胚胎癌细胞系P19进行传代培养,将细胞分为正常对照组(Con组)、添加反式维甲酸组(RA组)、添加GSK-3β特异性抑制剂SB216763组(SB组)和添加SB216763+RA组(SB+RA组)四组。采用免疫荧光染色、RT-PCR和Western blotting法分别检测细胞中βcatenin、Oct4及C-myc的表达状况;通过BrdU标记与MTT法检测细胞的增殖。结果     加入SB216763可促进P19细胞β-catenin入核,同时促进Oct4和C-myc基因的表达,阻断RA诱导的细胞分化,并可明显促进细胞增殖。结论    激活Wnt/β-catenin信号通路可促进胚胎癌细胞P19的增殖并抑制分化,其作用机制可能是通过上调C-myc基因表达来实现的。

关键词: 胚胎癌细胞;β-catenin;C-myc;增殖

Abstract:

Objective     To explore the role and mechanism of  the Wnt/β-catenin pathway on proliferation of embryonic carcinoma cells(EC). Methods     P19 cells, a mouse EC cell line, were divided into four groups: the normal control group(con), the RA group (medium with RA), the SB group(medium with GSK3β inhibitor SB216763), and SB+RA group (medium with both SB216763 and RA). At different culture times, immunofluorescence staining, RT-PCR and Western blotting were employed to observe expression of β-catenin, Oct4 and C-myc. BrdU labeling and MTT were used to measure cell proliferation. Results      The addition of SB216763 improved the nuclear location of β-catenin, high expression of Oct4 and C-myc, blocked P19 cell differentiation induced by RA, and dramatically promoted P19 cell proliferation. Conclusion      Activation of the Wnt/β-catenin pathway could promote proliferation and inhibit differentiation of mouse EC cells by upregulating C-myc expression.

Key words: Embryonic carcinoma cells; β-catenin; C-myc; Proliferation

中图分类号: 

  • R737.21
[1] 李毅1,张帅2,殷钢3,史琨1,邴鲁军1,郝晶1. Wnt/β-catenin信号通路在成人睾丸支持细胞增殖中的作用及其机制[J]. 山东大学学报(医学版), 2012, 50(3): 55-61.
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