不同家禽细胞Wnt/β-catenin信号通路基础活化状态的比较  

Comparison of basal activation status of Wnt/β-catenin signaling pathway in different poultry cells

作　　者：乔丹丹 程晓薇[2] 何倩 秦爱建 钱琨[1,2,3,4] QIAO Dan-dan;CHENG Xiao-wei;HE Qian;QIN Ai-jian;QIAN Kun(Key Laboratory for Avian Preventive Medicine,Ministry of Education,Yangzhou University,Yangzhou 225009,China;Key Laboratory of Jiangsu Preventive Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou University,Yangzhou 225009,China;Joint International Research Laboratory of Agriculture&Agri-Product Safety,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学教育部禽类预防医学重点实验室,江苏扬州225009 [2]扬州大学江苏省动物预防医学重点实验室,江苏扬州225009 [3]扬州大学江苏高校动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009 [4]扬州大学教育部农业与农产品安全国际合作联合实验室,江苏扬州225009

出　　处：《中国兽医科学》2019年第11期1434-1439,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目（31772734）;国家自然科学基金国际（地区）间重大项目（31761133002）;扬州大学研究生科研创新计划项目（XKYCX19-129）;江苏省优势学科项目

摘　　要：了解家禽细胞静息状态下Wnt/β-catenin信号通路的基础活化状态,为研究家禽Wnt信号通路提供细胞模型。利用激光共聚焦显微镜、绝对荧光定量PCR和Western-blot方法检测鸡胚成纤维细胞(CEF)、鸡胚肾细胞(CEK)、鸡成纤维永生化细胞系(DF-1)和鸡肝癌细胞系(LMH)中β-catenin蛋白的定位情况以及表达情况。同时使用双荧光素酶报告基因检测不同细胞加入激活剂后Wnt信号通路的活化情况。结果显示,CEF细胞中β-catenin基因和蛋白表达水平较低,主要位于细胞膜上,且对信号通路激活剂反应最为明显。DF-1细胞与LMH细胞中β-catenin的mRNA水平与蛋白水平均高于正常家禽细胞。结果表明,CEF细胞Wnt/β-catenin信号通路基础活化状态较低,是研究家禽Wnt相关信号通路的良好细胞模型。To understand the basal activation status of Wnt/β-catenin signaling pathway in poultry resting cells,and to provide a cell model for studying Wnt signaling pathway in poultry,the localization and expression ofβ-catenin in chicken embryo fibroblasts(CEF),chicken embryonic kidney cells(CEK),chicken fibroblast immortalized cell line(DF-1)and chicken hepatoma cell line(LMH)were detected by laser confocal microscopy,absolute fluorescent quantitative PCR and Western-blot methods.The activation of Wnt signaling pathway was detected by dual luciferase activities after addition of activator in different cells.The results showed that the gene and protein expression levels ofβ-catenin in the CEF cells were very low,and it mainly located on the cell membrane.But CEF cells were most responsive to signaling pathway activators.The expression level ofβ-catenin in DF-1 and LMH cells were higher than that of other poultry cells.In conclusion,the basal activation status of Wnt/β-catenin signaling pathway in CEF cells was low,which provides a good cell model for further study Wnt signaling pathway in poultry.

关 键 词：家禽细胞 WNT Β-连环蛋白 信号通路 基础活化状态 

分 类 号：S852.6[农业科学—基础兽医学]