摘要
目的比较OCT4异构体(OCT4A、OCT4B、OCT4B1)及其调控因子在人胚胎干细胞(hESC)和人间充质干细胞(hMSC)中的表达。方法利用RT-PCR、免疫荧光染色、流式细胞分析及体内/外分化实验,鉴定hESC及hMSC的生物学特性;应用real-time PCR、Western blot和流式细胞分析比较OCT4异构体及其转录因子NANOG,SOX2和mRNA结合蛋白LIN28在hESC及hMSC中的表达水平。结果 OCT4异构体mRNA在hESC和hMSC中均有表达,在hESC中的表达显著高于在hMSC中,并以OCT4A的差别最为显著(P<0.01);在蛋白水平,hESC表达OCT4A和OCT4B-256aa,hMSC不表达OCT4异构体蛋白。hESC高表达OCT4的调控因子NANOG、SOX2和LIN28;hMSC低表达SOX2,不表达NANOG和LIN28。结论 NANOG、SOX2和LIN28调控OCT4的表达,OCT4异构体在hESC和hMSC中的表达差异提示其可能是不同发育阶段干细胞自我更新和分化潜能等方面差别的主要因素之一。
Objective To compare the expression of OCT4 isoforms in human embryonic stem cells（ hESCs） and in human mesenchymal stem cells（ hMSCs）. Methods The characterization of hESCs and hMSCs were performed by RT-PCR,immunofluorescence staining,flow cytometry and in vivo / vitro differentiation assays. The expressions of OCT4 isoforms and regulators that control their transcription（NANOG,SOX2）and translation（LIN28）were quantified by real-time PCR,flow cytometry and Western blot. Results Three OCT4 isoforms of mRNA were ex-pressed in both hESC and hMSC. However,their expression was significantly higher in hESC than that in hMSC, especially for OCT4A（P 〈0. 01）. At protein level,OCT4A and OCT4B-265aa were translated in hESC while no OCT4 protein was be detected in hMSC. NANOG,SOX2 and LIN28 were highly translated in hESCs while hMSCs were weak positive for SOX2 expression and negative for NANOG and LIN28. Conclusions NANOG,SOX2 and LIN28 regulate the expression of OCT4. The different expression profile of OCT4 isoforms in hESC and hMSC indi-cates that OCT4 may be one of the important factors resulting in the differences of self-renewal and differentiation potentials of stem cells at different developmental stages.
出处
《基础医学与临床》
CSCD
北大核心
2014年第1期41-46,共6页
Basic and Clinical Medicine
基金
国家自然科学基金(31201102
30871433
31071305)
关键词
人胚胎干细胞
间充质干细胞
OCT4
选择性剪切
human embryonic stem cells
human mesenchymal stem cells
OCT4
alternative splicing
