基础医学与临床 ›› 2014, Vol. 34 ›› Issue (6): 723-728.

• 研究论文 •    下一篇

FGFR2异构体及相关剪接因子在人胚胎干细胞和成体干细胞中的表达

傅歆,谷聪敏,严笠,肖苒   

  1. 中国医学科学院整形外科医院
  • 收稿日期:2013-12-23 修回日期:2014-03-14 出版日期:2014-06-05 发布日期:2014-05-26
  • 通讯作者: 肖苒 E-mail:xiaoran@pumc.edu.cn
  • 基金资助:
    RNA选择性剪接在骨髓间充质干细胞定向分化中的调控研究;视黄酸对ALX3的基因调控作用和分子机制研究;转录因子KLF15在骨髓间充质干细胞定向分化中的调控作用研究

Expressions of FGFR2 isoforms and the relevant alternative splicing factors in human embryonic stem cells and adult stem cells

  • Received:2013-12-23 Revised:2014-03-14 Online:2014-06-05 Published:2014-05-26
  • Contact: Ran XIAO E-mail:xiaoran@pumc.edu.cn

摘要: 目的 比较FGFR2异构体及其剪接因子在人胚胎干细胞(hESCs)及其分化的拟胚体(EBs),以及骨髓间充质干细胞(BMSCs)、脂肪间充质干细胞(ADSCs)和毛囊干细胞(HFSCs)等不同组织来源成体干细胞(ASCs)中的表达。方法 用RT-PCR、免疫荧光染色和流式细胞分析,鉴定hESCs、EBs和ASCs的生物学特性;用real-time PCR比较FGFR2异构体IIIb,IIIc及相关剪接因子的表达。结果 FGFR2的选择性剪接在hESCs,EBs和ASCs中均以IIIc为主。hESCs向EBs分化后IIIb和IIIc的表达及IIIb/IIIc比值均升高,EBs中抑制IIIc表达的剪接因子FOX2和抑制IIIb表达的剪接因子HnRNPA1分别在分化第7天和分化第14天表达升高(P<0.05)。FGFR2异构体在hESCs与HFSCs中的表达水平显著高于在BMSCs和ADSCs中(P<0.01),在BMSCs中的表达水平显著高于在ADSCs中(P<0.05)。多种FGFR2相关剪接因子在hESCs和HFSCs中的表达水平显著高于在BMSCs和ADSCs中(P<0.05)。结论FGFR2异构体及其相关剪接因子在不同分化阶段和组织来源的干细胞中有不同的表达谱。

关键词: 人胚胎干细胞, 成体干细胞, FGFR2, 选择性剪接

Abstract: Objective To compare the expressions of FGFR2 isoforms and the relevant alternative splicing factors in human embryonic stem cells (hESCs) and their differentiated embryoid bodies (EBs), and human adult stem cells (ASCs) derived from different tissue sources, including bone marrow derived mesenchymal stem cells (BMSCs), adipose derived mesenchymal stem cell (ADSCs) and hair follicle stem cells (HFSCs). Methods The characteristics of hESCs, EBs and ASCs were confirmed by RT-PCR, immunofluorescence staining and flow cytometry analysis. The expressions of FGFR2 isoforms and the relevant splicing factors were quantified by real-time PCR. Results FGFR2 IIIc is the dominant isoform in all tested cells. Upon differentiation, the expressions of FGFR2 isoforms, as well as the IIIb/IIIc ratio in EBs were significantly increased (P<0.05). The expression of splicing factors that inhibited IIIc expression (FOX2) and inhibited IIIb expression (HnRNPA1) in EBs was increased after differentiation for 7 days and 14 days respectively (P<0.05). The expressions of FGFR2 isoforms in hESCs and HFSCs were significantly higher than in BMSCs and ADSCs, and significantly higher in BMSCs than in ADSCs (p<0.05). Expressions of multiple FGFR2 relevant splicing factors were significantly higher in hESCs and HFSCs than in BMSCs and ADSCs. Conclusion The expression profiles of FGFR2 isoforms and the relevant splicing factors in stem cells at different developmental stages and from different tissue sources are different.

Key words: human embryonic stem cells, adult stem cells, FGFR2, alternative splicing

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