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首页> 外文期刊>Cell biology international. >Inhibition of transcription factor T‐cell factor 3 (TCF3) using the oligodeoxynucleotide strategy increases embryonic stem cell stemness: possible application in regenerative medicine
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Inhibition of transcription factor T‐cell factor 3 (TCF3) using the oligodeoxynucleotide strategy increases embryonic stem cell stemness: possible application in regenerative medicine

机译：使用寡脱氧核苷酸核苷酸官果的转录因子T细胞因子3（TCF3）的抑制增加了胚胎干细胞茎：可能在再生医学中的应用

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Abstract The transcription factor T‐cell factor 3 (TCF3), one component of the Wnt pathway, is known as a cell‐intrinsic inhibitor of many pluripotency genes in embryonic stem cells (ESCs) that influences the balance between pluripotency and differentiation. In this study, the effects of inhibition of TCF3 transcription factor on the stemness of mouse ESCs (mESCs) were investigated using the decoy oligodeoxynucleotides (ODNs) strategy. The TCF3 decoy and its scramble ODNs were designed and synthesized. The interaction specificity of the TCF3 decoy with the TCF3 transcription factor was evaluated by the electrophoretic mobility shift assay. Subcellular localization was carried out using fluorescence and confocal microscopy. Self‐renewal and pluripotency of mESCs were analyzed by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide (MTT), cell cycle and apoptosis, alkaline phosphatase (ALP), embryoid body (EB) formation, and real‐time assays. All experiments were performed in triplicate. The results showed that knockdown of TCF3 by decoy ODNs transfection in mESCs led to an increase in the cell proliferation, ALP enzyme activity, and master regulatory stemness genes and a decrease in the number and diameter of EBs. These results supported TCF3 as a potential target to maintain the pluripotency and self‐renewal capacity of mESCs. Knockdown of the TCF3 transcription factor using decoy ODNs can be a promising method to maintain the stemness of stem cells in regenerative medicine and cell therapy researches.
机译：摘要，转录因子T细胞因子3（TCF3），WNT途径的一种组分称为胚胎干细胞（ESC）中许多多能性基因的细胞内在抑制剂，其影响多能性和分化之间的平衡。在该研究中，使用诱饵寡核苷核苷酸（ODN）策略研究了抑制TCF3转录因子对小鼠ESC（MESCS）茎的影响。设计和合成了TCF3诱饵及其争夺ODN。通过电泳迁移率移位测定评估TCF3诱饵与TCF3转录因子的相互作用特异性。使用荧光和共聚焦显微镜进行亚细胞定位。通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴铵（MTT），细胞周期和凋亡，碱性磷酸酶（ALP），胚胎体（EB）分析自我更新和多能性）形成和实时测定。所有实验均以三份进行。结果表明，MESCS中诱饵ODN转染的TCF3的敲低导致细胞增殖，ALP酶活性和母体调节性茎秆基因的增加和EBS数量和直径的降低。这些结果支持TCF3作为维持​​MESCS多能性和自我更新能力的潜在目标。使用诱饵ODN的TCF3转录因子的敲低可以是保持干细胞在再生医学和细胞疗法研究中的干细胞茎的方法。

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