吉林大学学报(医学版)

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T细胞因子3基因沉默对小鼠睾丸胚胎癌细胞Oct4表达的影响

林桂淼1,文娟2,陈强1,王晓梅1   

  1. (1.   深圳大学医学院泌尿生殖研究所和合成生物学工程实验室,广州 深圳 518066;2.新疆医科大学基础医学院病理生理学教研室,新疆 乌鲁木齐 830011)
  • 收稿日期:2013-07-15 出版日期:2014-01-28 发布日期:2014-01-28
  • 通讯作者: 王晓梅 E-mail:(Tel:0755-86671908, E-mail: xmwang@szu.edu.cn)
  • 作者简介:林桂淼(1980-),女,福建省永安市人,讲师,医学博士,主要从事肿瘤基因治疗和分子机制研究。
  • 基金资助:

    国家自然科学基金资助课题(81301318);广东省科技厅科技计划项目资助课题(2011B031800351);广东省深圳市科技创新委员会基础研究项目资助课题(JCYJ20120613170218654)

Effect of T cell factor 3 gene silencing on expression of Oct4 in mouse embryonic carcinoma cells

LIN Gui-miao1,WEN Juan2,CHEN Qiang1,WANG Xiao-mei1   

  1. (1. Laboratory of Synthetic Biology Engineering,Research Institute of Uropoiesis and Reproduction,School of Medicine,Shenzhen University,Shenzhen 518066,China;2. Department of Pathophysiology,College of Basic Medical Sciences,Xinjiang Medical University,Urumqi 830011,China)
  • Received:2013-07-15 Online:2014-01-28 Published:2014-01-28

摘要:

目的:探讨T细胞因子3 (TCF3)基因沉默对小鼠睾丸胚胎癌F9细胞干性基因Oct4表达的影响,阐明TCF3对F9细胞中Oct4表达调控的作用机制。方法:采用RNAi技术将TCF3基因沉默作为TCF3干涉组,同时设立空白对照组和阴性对照组。采用RT-PCR和Western blotting方法观察各组细胞TCF3和Oct4 mRNA和蛋白表达水平,细胞免疫荧光法观察F9细胞中Oct4的表达。结果:Oct4在F9细胞核中高度表达。与空白对照和阴性对照组比较,TCF3干涉组TCF3 mRNA和蛋白表达水平显著下调,差异有统计学意义(P<0.01);Oct4 mRNA和蛋白表达水平显著升高,差异有统计学意义(P<0.01);而空白对照组与阴性对照组间比较差异无统计学意义(P>0.05)。结论: TCF3基因干涉可以促进F9细胞中Oct4的转录和蛋白表达,提示TCF3可以负向调控F9细胞中Oct4的表达,是Oct4的抑制因子之一。

关键词: 胚胎肿瘤, T细胞因子3, 八聚体4, 基因沉默

Abstract:

To investigate the effect of T cell factor 3(TCF3) gene silencing on the expression of Oct4 and to explore the mechanism of Oct4 expression regulation by TCF3 in mouse embryonic carcinoma F9 cells. Methods The TCF3 gene in F9 cells was silenced by RNAi technology and used as TCF3 silencing group;meanwhile blank control group (mock) and negative control group (scramble) were set up. The mRNA and protein expression levels of TCF3 and Oct4 in F9 cells in various groups were detected by RT-PCR and Western blotting methods.The expression of Oct4 in F9 cells was detected by immunofluorescence assay.Results Oct4 was highly expressed in nucleus of F9 cells.Compared with mock group and scramble group,the mRNA and protein expression levels of TCF3 in TCF3 silencing group were significantly decreased(P<0.01),while the mRNA and protein expression levels of Oct4 were significantly increased(P<0.01).There was no significant difference between mock group and scramble group(P>0.05).Conclusion Gene silencing of TCF3 in F9 cells can obviously promote the transcription and expression of Oct4,which indicates that TCF3 can negatively regulate the expression of Oct4 in F9 cells as one of the inhibitors.

Key words: embryonic neoplasms, T cell factor 3, octamer 4, gene silencing

中图分类号: 

  • Q78