【摘要】：目的比較OCT4異構(gòu)體(OCT4A、OCT4B、OCT4B1)及其調(diào)控因子在人胚胎干細(xì)胞(hESC)和人間充質(zhì)干細(xì)胞(hMSC)中的表達(dá)。方法利用RT-PCR、免疫熒光染色、流式細(xì)胞分析及體內(nèi)/外分化實(shí)驗(yàn),鑒定hESC及hMSC的生物學(xué)特性;應(yīng)用real-time PCR、Western blot和流式細(xì)胞分析比較OCT4異構(gòu)體及其轉(zhuǎn)錄因子NANOG,SOX2和mRNA結(jié)合蛋白LIN28在hESC及hMSC中的表達(dá)水平。結(jié)果 OCT4異構(gòu)體mRNA在hESC和hMSC中均有表達(dá),在hESC中的表達(dá)顯著高于在hMSC中,并以O(shè)CT4A的差別最為顯著(P0.01);在蛋白水平,hESC表達(dá)OCT4A和OCT4B-256aa,hMSC不表達(dá)OCT4異構(gòu)體蛋白。hESC高表達(dá)OCT4的調(diào)控因子NANOG、SOX2和LIN28;hMSC低表達(dá)SOX2,不表達(dá)NANOG和LIN28。結(jié)論 NANOG、SOX2和LIN28調(diào)控OCT4的表達(dá),OCT4異構(gòu)體在hESC和hMSC中的表達(dá)差異提示其可能是不同發(fā)育階段干細(xì)胞自我更新和分化潛能等方面差別的主要因素之一。
[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.
【作者單位】： 中國(guó)醫(yī)學(xué)科學(xué)院整形外科醫(yī)院研究中心;
【基金】：國(guó)家自然科學(xué)基金(31201102,30871433,31071305)
【分類號(hào)】：R329

【參考文獻(xiàn)】

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