Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland.
Sci Rep. 2018 Apr 3;8(1):5425. doi: 10.1038/s41598-018-23794-5.
Recent studies have shown that the transcription factor Foxn1, which is expressed in keratinocytes, is involved in the skin wound healing process, yet how Foxn1 functions remains largely unknown. Our latest data indicate that Foxn1 drives skin healing via engagement in re-epithelization and the epithelial-mesenchymal transition (EMT) process. In the present study, 2D-DIGE proteomic profiling analysis of in vitro cultured keratinocytes transfected with adenoviral vector carrying Foxn1-GFP or GFP alone (control) revealed forty proteins with differential abundance between the compared groups. Among the proteins with Foxn1-dependent expression, several enable adaptation to hypoxia. Subsequent experiments revealed that hypoxic conditions (1% O) stimulate endogenous and exogenous (transfected Ad-Foxn1) Foxn1 expression in cultured keratinocytes. A proteomics analysis also identified proteins that can act as a factors controlling the balance between cell proliferation, differentiation and apoptosis in response to Foxn1. We also showed that in C57BL/6 keratinocytes, the stimulation of Foxn1 by hypoxia is accompanied by increases in Mmp-9 expression. These data corroborate the detected co-localization of Foxn1 and Mmp-9 expression in vivo in post-wounding skin samples of Foxn1::Egfp transgenic mice. Together, our data indicate that Foxn1 orchestrates cellular changes in keratinocytes in both physiological (self-renewal) and pathological (skin wound healing) contexts.
最近的研究表明，转录因子 Foxn1 在角质细胞中表达，参与皮肤伤口愈合过程，但 Foxn1 的功能仍知之甚少。我们的最新数据表明，Foxn1 通过参与再上皮化和上皮-间充质转化（EMT）过程来驱动皮肤愈合。在本研究中，对转染携带 Foxn1-GFP 或 GFP 载体的腺病毒的体外培养角质细胞进行 2D-DIGE 蛋白质组学分析，显示比较组之间有四十种蛋白丰度存在差异。在 Foxn1 依赖性表达的蛋白中，有几种能够适应缺氧。随后的实验表明，缺氧条件（1% O）刺激培养角质细胞内内源性和外源性（转染的 Ad-Foxn1）Foxn1 表达。蛋白质组学分析还鉴定出一些能够作为控制细胞增殖、分化和凋亡平衡的因子的蛋白，以响应 Foxn1。我们还表明，在 C57BL/6 角质细胞中，缺氧刺激 Foxn1 会伴随着 Mmp-9 表达的增加。这些数据证实了在 Foxn1::Egfp 转基因小鼠伤口后皮肤样本中体内检测到的 Foxn1 和 Mmp-9 表达的共定位。总之，我们的数据表明，Foxn1 在生理（自我更新）和病理（皮肤伤口愈合）环境中协调角质细胞的细胞变化。
