转录因子NKX2.1促进诱导多能干细胞向肺干细胞的分化  

Transcription factor NKX2.1 promotes differentiation of induced pluripotent stem cells into lung stem cells

作　　者：邓丽 刘洋 王慧[1] 杨秋[1] 董明清 Deng Li;Liu Yang;Wang Hui;Yang Qiu;Dong Mingqing(Center for Medicine Research and Translation,Chengdu Fifth People’s Hospital,Chengdu 611130,Sichuan Province,China)

机构地区：[1]成都市第五人民医院医学研究与转化中心,四川省成都市611130

出　　处：《中国组织工程研究》2025年第36期7790-7796,共7页Chinese Journal of Tissue Engineering Research

基　　金：四川省科技厅自然科学基金项目(2023NSFSC0531),项目负责人:董明清;成都市高水平临床重点专科建设项目(KYJJ2021-27),项目负责人:董明清;成都中医药大学“杏林学者”医院专项(XJ2023010001),项目负责人:邓丽。

摘　　要：目的:探究NKX2.1对诱导多能干细胞向肺干细胞方向分化的影响。方法:体外培养人诱导多能干细胞,采用实时荧光定量PCR和免疫荧光检测多能干细胞特异基因的表达;通过瞬时转染向人诱导多能干细胞中过表达NKX2.1,再诱导其向肺干细胞方向分化,诱导分化7 d采用实时荧光定量PCR、免疫荧光检测FoxA2、SOX9和P63的表达,诱导分化13 d采用免疫荧光检测肺泡细胞标记分子SPB和SPC的表达。结果与结论:(1)诱导多能干细胞紧密集结呈典型克隆样生长,显著高表达干细胞特异表达基因OCT-4、SOX2和NANOG;(2)与未转染对照组相比,过表达NKX2.1组人诱导多能干细胞中NKX2.1表达显著升高(P<0.000 1);(3)诱导分化7 d,与未转染对照组相比,过表达NKX2.1组肺干细胞相关标记物FoxA2、SOX9和P63的表达显著升高(P<0.000 1);(4)诱导分化13 d,与未转染对照组相比,过表达NKX2.1组肺泡细胞标记分子SPB和SPC荧光强度显著增加。结果表明NKX2.1可以促进诱导多能干细胞向肺干细胞方向分化。BACKGROUND:Enhancing the differentiation of induced pluripotent stem cells into lung stem cells is crucial for repairing lung injuries.NKX2.1 is the earliest marker of lung epithelial differentiation and plays a significant regulatory role in lung development.However,the impact of its expression on the differentiation of induced pluripotent stem cells into lung stem cells remains inadequately understood.OBJECTIVE:To investigate the effect of NKX2.1 on the differentiation of induced pluripotent stem cells into lung stem cells.METHODS:Induced pluripotent stem cells were cultured in vitro.The expression of specific pluripotent stem cell genes was assessed using real-time fluorescence quantitative PCR.NKX2.1 was overexpressed in induced pluripotent stem cells,which were then induced to differentiate into lung stem cells.The expression of FoxA2,SOX9,and P63 was determined via quantitative PCR and immunofluorescence on day 7 of induction of differentiation.The expression of the alveolar marker SPB and SPC was evaluated through immunofluorescence staining on day 7 of induction of differentiation.RESULTS AND CONCLUSION:(1)Induced pluripotent stem cells in vitro were tightly packed and showed typical clonoid growth and significantly expressed stem cell-specific genes OCT-4,SOX2,and NANOG.(2)Compared with the non-transfected control group,the expression of NKX2.1 in human induced pluripotent stem cells was significantly increased in the NKX2.1 overexpression group(P<0.0001).(3)Seven days after induction of differentiation,compared with the non-transfected control group,the expression of lung stem cell-related markers FoxA2,SOX9,and P63 was significantly increased in the NKX2.1 overexpression group(P<0.0001).(4)Thirteen days after induction of differentiation,compared with the non-transfected control group,the fluorescence intensity of alveolar cell marker molecules SPB and SPC increased significantly in the overexpression NKX2.1 group.The results show that NKX2.1 can promote the differentiation of induced pluripotent stem

关 键 词：诱导多能干细胞 肺干细胞 NKX2.1 定型内胚层 前肠内胚层 肺损伤 工程化干细胞 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R563