OCT4异构体在人胚胎干细胞和间充质干细胞中的表达
发布时间：2018-08-21 20:47
【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(P0.01);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。
[Abstract]:Objective to compare the expression of OCT4 isomer (OCT4AnOCT4B1) and its regulatory factors in human embryonic stem cell (hESC) and human mesenchymal stem cell (hMSC). Methods the biological characteristics of hESC and hMSC were identified by RT-PCR, immunofluorescence staining, flow cytometry and in vitro / in vivo differentiation assay. The expression levels of OCT4 isomer and its transcription factor NANOGN SOX2 and mRNA binding protein LIN28 in hESC and hMSC were compared by real-time PCR Western blot and flow cytometry. Results OCT4 isomer mRNA was expressed in both hESC and hMSC, and the expression in hESC was significantly higher than that in hMSC. The expression of OCT4A and OCT4B-256aahMSC did not express OCT4 isomer protein HESC and the regulatory factors of high expression of OCT4, NANOGG SSOX2 and LIN28hMSC, but not NANOG and LIN28hMSC. Conclusion the difference in the expression of OCT4 isomers in hESC and hMSC suggests that NANOGN SOX2 and LIN28 may be one of the main factors of the difference in self-renewal and differentiation potential of stem cells in different developmental stages.
【作者单位】： 中国医学科学院整形外科医院研究中心;
【基金】：国家自然科学基金(31201102,30871433,31071305)
【分类号】：R329
[Abstract]:Objective to compare the expression of OCT4 isomer (OCT4AnOCT4B1) and its regulatory factors in human embryonic stem cell (hESC) and human mesenchymal stem cell (hMSC). Methods the biological characteristics of hESC and hMSC were identified by RT-PCR, immunofluorescence staining, flow cytometry and in vitro / in vivo differentiation assay. The expression levels of OCT4 isomer and its transcription factor NANOGN SOX2 and mRNA binding protein LIN28 in hESC and hMSC were compared by real-time PCR Western blot and flow cytometry. Results OCT4 isomer mRNA was expressed in both hESC and hMSC, and the expression in hESC was significantly higher than that in hMSC. The expression of OCT4A and OCT4B-256aahMSC did not express OCT4 isomer protein HESC and the regulatory factors of high expression of OCT4, NANOGG SSOX2 and LIN28hMSC, but not NANOG and LIN28hMSC. Conclusion the difference in the expression of OCT4 isomers in hESC and hMSC suggests that NANOGN SOX2 and LIN28 may be one of the main factors of the difference in self-renewal and differentiation potential of stem cells in different developmental stages.
【作者单位】： 中国医学科学院整形外科医院研究中心;
【基金】：国家自然科学基金(31201102,30871433,31071305)
【分类号】：R329
【参考文献】
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