p53-NF-κB Crosstalk in Febrile Temperature-Treated Human Umbilical Cord-Derived Mesenchymal Stem Cells
p53-NF-κB Crosstalk in Febrile Temperature-Treated Human Umbilical Cord-Derived Mesenchymal Stem Cells
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DOI：
10.1089/scd.2018.0115
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发表时间：
2018-11-15
影响因子：
4
通讯作者：
Ta, Malancha
中科院分区：
文献类型：
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作者：
Goyal, Umesh;Ta, Malancha
间充质干细胞（MSC）由于其抗炎、营养和免疫调节作用而成功地用于免疫和炎症背景中的治疗应用。然而，尽管间充质干细胞有潜力为各种破坏性疾病提供再生治疗，但移植间充质干细胞的大量细胞死亡仍然是一个需要克服的障碍。间充质干细胞与炎症之间的关系是多因素的，难以理解。发烧是发炎微环境的关键组成部分。此外，MSC来源的选择可能是决定移植细胞在应激条件下的命运的关键。在这里，我们研究了沃顿氏胶质间充质干细胞（WJ-MSCs）的温度敏感性升高的温度在生理发热范围。我们探讨了发热范围温度对WJ-MSCs的形态、活力、增殖动力学和细胞周期状态的影响。WJ-MSCs在40 ℃下呈扁平状，我们使用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四唑溴化物（MTT）进行的增殖动力学研究和凋亡测定的数据显示，与对照培养物相比，WJ-MSCs在40 ℃下的增殖和活力降低。也有G 0/G1细胞周期停滞，这进一步证实了信使RNA（mRNA）水平的基因特异性的细胞周期的不同阶段。在评估p53状态时，我们观察到暴露于40 ℃的WJ-MSCs中p53蛋白表达及其核定位增加。其下游效应子p21也被上调。此外，这种温度诱导的p53诱导在NF-B途径抑制剂、吡咯烷二硫代氨基甲酸酯（PDTC）或靶向p65的核酸内切酶制备的小干扰RNA（esiRNA）存在下暴露于40 ℃时被抑制。高温暴露不影响WJ-MSCs的衰老状态。在40 ° C下的MSC特异性表面抗原谱与对照WJ-MSC相似。我们的研究结果表明，在发热温度应激条件下，WJ-MSCs表现出G 0/G1细胞周期停滞和活细胞计数减少，同时保留其基本特征，与p53和NF-B通路的潜在相互作用。
Mesenchymal stem cells (MSCs) are successful for their therapeutic application in immune and inflammatory contexts due to their anti-inflammatory, trophic, and immunomodulatory roles. However, though MSCs have the potential to provide regenerative treatment toward a wide range of devastating diseases, massive cell death of transplanted MSCs remains an obstacle to overcome. The relation between MSCs and inflammation is multifactorial and challenging to comprehend. Fever is a critical component of the inflamed microenvironment. Also, the choice of MSC source could be critical in determining the fate of transplanted cells under stress conditions. Here we investigated the thermosensitivity of Wharton's jelly MSCs (WJ-MSCs) to elevated temperature in the physiological fever range. We explored the effect of febrile range temperature on morphology, viability, proliferation kinetics, and cell cycle status of WJ-MSCs. WJ-MSCs adopted a flattened morphology at 40 degrees C, and our data from proliferation kinetics study using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and apoptosis assays showed that WJ-MSCs had reduced proliferation and viability at 40 degrees C compared with control cultures. There was also a G0/G1 cell cycle arrest, which was further confirmed by messenger RNA (mRNA) levels of genes specific for different stages of cell cycle. On evaluating p53 status, we observed an increase in p53 protein expression and its nuclear localization in WJ-MSCs exposed to 40 degrees C. Its downstream effector p21 too was upregulated. Moreover, this temperature-induced p53 induction was inhibited on exposure to 40 degrees C in the presence of NF-B pathway inhibitor, pyrrolidinedithiocarbamate (PDTC) or endonuclease-prepared small interfering RNA (esiRNA) targeting p65. Febrile temperature exposure did not affect the senescence status of WJ-MSCs. The MSC-specific surface antigen profile at 40 degrees C was similar to control WJ-MSCs. Our findings suggest that under febrile temperature stress conditions, WJ-MSCs exhibit G0/G1 cell cycle arrest and reduction in viable cell count, while retaining their basic characteristics, with an underlying interplay of p53 and NF-B pathway.