Huang Liang-Liang, Deng Jun-Hong, Xie Jing-Xuan, Lin Zi-Bin, Jiang Hui, Ouyang Bin, Liu Jian-Ming, Wei Yan-Ni, Cai Zhou-da
Department of Andrology, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, Guangzhou 510180, China.
Department of Urology, Peking University Third Hospital, Andrology, Beijing 100191, China.
Stem Cells Int. 2021 Mar 27;2021:6633111. doi: 10.1155/2021/6633111. eCollection 2021.
Urinary stem cells (USCs) have the capacity for unlimited growth and are promising tools for the investigations of cell differentiation and urinary regeneration. However, the limited life span significantly restricts their usefulness. This study is aimed at exploring the effect of integrin-linked kinase (ILK) on the smooth muscle cells (SMCs) differentiation of the dog USCs and investigating its molecular mechanism.
An immortalized USCs cell line with the molecular markers and biological functions was prepared. After successfully inducing the differentiation of USCs into SMCs, the expression level of the unique key factor and its mechanisms in this process was determined through real-time polymerase chain reaction, Western blot, or Immunofluorescence staining.
We found that high cell density promoted USCs differentiation SMCs, and ILK was necessary for USCs differentiation into SMCs. Knocking down ILK decreased the expression of SMCs specific-marker, while using a selective ILK agonist increased the expression of SMCs specific-marker. Furthermore, ILK regulated SMCs differentiation in part through the activation of NF-B pathway in USCs. A NF-B activity assay showed overexpression of ILK could significantly upregulate NF-B p50 expression, and NF-B p50 acts as downstream signal molecular of ILK.
High cell density induces the differentiation of USCs into SMCs, and ILK is a key regulator of myogenesis. Furthermore, NF-B signaling pathway might play a crucial role in this process.
尿源干细胞（USCs）具有无限增殖能力，是细胞分化和尿再生研究的有前景的工具。然而，其有限的寿命显著限制了它们的用途。本研究旨在探讨整合素连接激酶（ILK）对犬USCs向平滑肌细胞（SMCs）分化的影响，并研究其分子机制。
制备具有分子标记和生物学功能的永生化USCs细胞系。在成功诱导USCs分化为SMCs后，通过实时聚合酶链反应、蛋白质免疫印迹或免疫荧光染色确定该过程中独特关键因子的表达水平及其机制。
我们发现高细胞密度促进USCs向SMCs分化，且ILK是USCs分化为SMCs所必需的。敲低ILK会降低SMCs特异性标志物的表达，而使用选择性ILK激动剂则会增加SMCs特异性标志物的表达。此外，ILK部分通过激活USCs中的NF-κB通路来调节SMCs分化。NF-κB活性检测显示ILK的过表达可显著上调NF-κB p50的表达，且NF-κB p50作为ILK的下游信号分子发挥作用。
高细胞密度诱导USCs分化为SMCs，且ILK是肌生成的关键调节因子。此外，NF-κB信号通路可能在此过程中起关键作用。
