摘要
目的比较FGFR2异构体及其剪接因子在人胚胎干细胞(h ESCs)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仪分析鉴定h ESCs、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h ESCs、EBs和ASCs中均以Ⅲc为主。h ESCs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天和第14天表达升高(P<0.05)。FGFR2异构体在h ESCs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h ESCs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。
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.
出处
《基础医学与临床》
CSCD
北大核心
2014年第6期723-728,共6页
Basic and Clinical Medicine
基金
国家自然科学基金(30871433
31201102
31071305)
