Zhang Jingru, Song Nan, Jia Xinshan, Sun Shanliang, Ma Xiaobo, Wang Enhua
Department of Pathology, College of Basic Medical Sciences, China Medical University, Shenyang 110001, China; Department of Pathology, the First Affiliated Hospital of China Medical University, Shenyang 110001, China.
Zhongguo Fei Ai Za Zhi. 2009 Feb 20;12(2):106-10. doi: 10.3779/j.issn.1009-3419.2009.02.03.
CAlthough there are increasing reports on localization of tracheal stem cells, the mechanism of proliferation and differentiation of tracheal stem cells remains unclear.
In this study, we developed an ex vivo model of murine tracheal epithelial injury and regeneration induced by 5-FU. The regeneration process of murine tracheal epithelium was observed and analyzed by morphological, immunofluorescence and microarray methods.
After treatment with 5-FU, the differentiated mature cells were dead and desquamated. Only a few cells remained in G0 phase and located on the basement membrane. After being put back in normal culture media, the cells became flat, cubic and restored as pseudostratified epithelium at last. These G0 phase cells were ABCG2 positive. We detected the differences of stem cell genes between normal tracheal epithelial cells and regenerated epithelial cells at 24 h and 48 h after injured by 5-FU using stem cell differentiation gene microarray. At 24 h, 8 genes were up-regulated and 31 genes were down-regulated. At 48 h, 5 genes were up-regulated and 42 genes were down-regulated.
The differently expressed genes were mainly associated with cell cycle regulation, intercellular junction, FGF, BMP, Notch and Wnt signaling pathways, which suggested these alterations might be closely associated with the proliferation and differentiation of tracheal stem cells.
尽管关于气管干细胞定位的报道日益增多,但气管干细胞增殖和分化的机制仍不清楚。
在本研究中,我们建立了5-氟尿嘧啶诱导的小鼠气管上皮损伤和再生的体外模型。通过形态学、免疫荧光和基因芯片方法观察和分析小鼠气管上皮的再生过程。
5-氟尿嘧啶处理后,分化成熟的细胞死亡并脱落。仅少数细胞处于G0期并位于基底膜上。放回正常培养基后,细胞变扁平、立方状,最终恢复为假复层上皮。这些G0期细胞ABCG2呈阳性。我们使用干细胞分化基因芯片检测了5-氟尿嘧啶损伤后24小时和48小时正常气管上皮细胞与再生上皮细胞之间干细胞基因的差异。在24小时时,8个基因上调,31个基因下调。在48小时时,5个基因上调,42个基因下调。
差异表达的基因主要与细胞周期调控、细胞间连接、FGF、BMP、Notch和Wnt信号通路相关,这表明这些改变可能与气管干细胞的增殖和分化密切相关。