Overexpression of FoxM1 promotes differentiation of bone marrow mesenchymal stem cells into alveolar type II cells through activating Wnt/β-catenin signalling
Overexpression of FoxM1 promotes differentiation of bone marrow mesenchymal stem cells into alveolar type II cells through activating Wnt/β-catenin signalling
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DOI：
10.1016/j.bbrc.2020.05.042
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发表时间：
2020-07-23
影响因子：
3.1
通讯作者：
Lin，Shan
中科院分区：
文献类型：
--
作者：
Zeng，Mian;Chen，Qingui;Lin，Shan
背景急性呼吸窘迫综合征（acute respiratory distress syndrome， ARDS）由于缺乏有效的治疗手段，已成为重症医学面临的严峻挑战。肺泡上皮损伤是ARDS的一个特征性特征，诱导间充质干细胞（mesenchymal stem cells, MSCs）分化为肺泡上皮细胞是治疗ARDS的一种很有前景的方法，但分化效率有待提高。本研究旨在探讨FoxM1过表达对MSCs向肺泡上皮细胞分化的影响。方法从小鼠骨髓中分离smscs，用携带FoxM1质粒的慢病毒转染。采用小气道上皮细胞生长培养基作为诱导间充质干细胞向肺泡上皮细胞分化的培养体系。分化效率主要通过定量逆转录聚合酶链反应和Western blot检测肺泡上皮细胞特异性标志物的表达水平来评估。为了研究Wnt/β-catenin信号是否参与调控机制，我们使用了一种特异性抑制剂XAV-939，并分别分析了核蛋白和细胞质蛋白。采用共免疫沉淀法检测FoxM1与β-catenin之间潜在的相互作用。结果过表达FoxM1可显著提高ⅱ型肺泡上皮细胞特异性标志物前表面活性蛋白C和表面活性蛋白B的表达水平，经XAV-939处理后可部分逆转，而ⅰ型肺泡上皮细胞特异性标志物水通道蛋白5的表达水平无显著变化。过表达FoxM1也增加了β-catenin的核易位及其转录活性。共免疫沉淀实验发现FoxM1与β-catenin之间存在直接相互作用。结论FoxM1过表达可部分通过激活Wnt/β-catenin信号通路提高MSCs向II型肺泡上皮细胞分化的效率。
BackgroundAcute respiratory distress syndrome (ARDS) becomes a serious challenge in critical care medicine due to the lack of effective therapy. As the damage of alveolar epithelium is a characteristic feature of ARDS, inducing mesenchymal stem cells (MSCs) to differentiate into alveolar epithelial cells turns out to be a promising therapy for ARDS, but the differentiation efficiency is yet to be improved. The study aimed to investigate the effect of overexpressing FoxM1 on MSCs’ differentiation into alveolar epithelial cells.MethodsMSCs were isolated from mouse bone marrow, followed by transfected with lentivirus carrying the FoxM1 plasmid. Small airway epithelial cell growth medium was used as a culture system for inducing MSCs’ differentiation into alveolar epithelial cells. Differentiation efficiency was assessed by detecting the expression levels of specific markers of alveolar epithelial cells mainly using quantitative reverse-transcription polymerase chain reaction and Western blot. To examine whether Wnt/β-catenin signalling was involved in the regulation mechanism, a specific inhibitor of the pathway XAV-939 was used and nuclear and cytoplasmic proteins were also analysed respectively. Co-immunoprecipitation was performed to examine the potential interaction between FoxM1 and β-catenin.ResultsOverexpressing FoxM1 statistically significantly increased the expression levels of specific markers of type II alveolar epithelial cells prosurfactant protein C and surfactant protein B, which was partially reversed by XAV-939 treatment, while the expression levels of specific marker of type I alveolar epithelial cells aquaporin 5 did not change significantly. Overexpressing FoxM1 also increased the nuclear translocation of β-catenin and its transcriptional activity. A direct interaction between FoxM1 and β-catenin was found in co-immunoprecipitation assay.ConclusionOverexpression of FoxM1 could improve the efficiency of MSCs’ differentiation into type II alveolar epithelial cells partly by activating Wnt/β-catenin signalling.