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首页> 外文期刊>Cell biology international. >Potential role of heat shock proteins in neural differentiation of murine embryonal carcinoma stem cells (P19).
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Potential role of heat shock proteins in neural differentiation of murine embryonal carcinoma stem cells (P19).

机译：热激蛋白在小鼠胚胎癌干细胞（P19）神经分化中的潜在作用。

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摘要

HSPs (heat shock proteins) have been recognized to maintain cellular homoeostasis during changes in microenvironment. The present study aimed to investigate the HSPs expression pattern in hierarchical neural differentiation stages from mouse embryonal carcinoma stem cells (P19) and its role in heat stressed exposed cells. For induction of HSPs, cells were heated at 42 degrees C for 30 min and recovered at 37 degrees C in different time points. For neural differentiation, EBs (embryoid bodies) were formed by plating P19 cells in bacterial dishes in the presence of 1 mM RA (retinoic acid) and 5% FBS (fetal bovine serum). Then, on the sixth day, EBs were trypsinized and plated in differentiation medium containing neurobasal medium, B27, N2 and 5% FBS and for an extra 4 days. The expression of HSPs and neural cell markers were evaluated by Western blot, flow cytometry and immunocytochemistry in different stages. Our results indicate that HSC (heat shock constant)70 and HSP60 expressions decreased following RA treatment, EB formation and in mature neural cells derived from heat-stressed single cells and not heat-treated EBs. While the level of HSP90 increased six times following maturation process, HSP25 was expressed constantly during neural differentiation; however, its level was enhanced with heat stress. Accordingly, heat shock 12 h before the initiation of differentiation did not affect the expression of neuroectodermal and neural markers, nestin and beta-tubulin III, respectively. However, both markers increased when heat shock was induced after treatment and when EBs were formed. In conclusion, our results raise the possibility that HSPs could regulate cell differentiation and proliferation under both physiological and pathological conditions.
机译：HSP（热激蛋白）已被公认在微环境变化过程中保持细胞的稳态。本研究旨在调查小鼠胚胎癌干细胞（P19）在神经分化阶段的HSPs表达模式及其在热应激暴露细胞中的作用。为了诱导HSP，将细胞在42℃下加热30分钟，并在不同时间点在37℃下恢复。对于神经分化，在存在1 mM RA（视黄酸）和5％FBS（胎牛血清）的情况下，通过在细菌培养皿中铺板P19细胞形成EB（胚状体）。然后，在第六天，用胰蛋白酶消化EB，并将其接种在含有神经基础培养基，B27，N2和5％FBS的分化培养基中，再培养4天。通过蛋白质印迹，流式细胞术和免疫细胞化学在不同阶段评估HSPs和神经细胞标志物的表达。我们的结果表明，在RA处理，EB形成以及源自热应激单细胞而非未经热处理的EB的成熟神经细胞中，HSC（热激常数）70和HSP60的表达降低。在成熟过程中，HSP90的水平增加了六倍，而HSP25在神经分化过程中不断表达。然而，其水平随着热应激而增加。因此，分化开始前12 h的热休克分别不影响神经外胚层和神经标志物Nestin和β-tubulinIII的表达。但是，当治疗后引起热激和形成EB时，两种标记物均增加。总之，我们的结果提高了HSPs在生理和病理条件下均可以调节细胞分化和增殖的可能性。

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