ITGB4BP对Wntβ-catenin信号通路活性的下调作用-第三军医大学学报.docVIP

ITGB4BP对Wnt/β-catenin信号通路活性的下调作用 赖庆梧，2 彭 旭2 谭江琳2 张小容 2胡小红2张 毅1罗高兴2吴 军2 (1华南理工大学生物科学与工程学院，广州 510006；2第三军医大学西南医院全军烧伤研究所，创伤、烧伤与复合伤国家重点实验室，疾病蛋白质组学重庆市市级重点实验室，重庆 400038) 摘要：目的 探讨β4整合素结合蛋白ITGB4BP对Wnt/β-catenin信号通路的影响。方法 将对照组空载体pCMV-3B质粒和实验组pCMV-ITGB4BP质粒分别转染至HEK293细胞中，用Western blot和激光共聚焦技术检测细胞中β-catenin含量的变化，用含TCF/LEF启动子的荧光素酶TOPFlash质粒和内参pRL-TK质粒共转染加有LiCl或未加LiCl的对照组和实验组的HEK293细胞，用荧光素酶活性分析法检测ITGB4BP转染后HEK293细胞TCF/LEF活性的变化。结果 ITGB4BP使HEK293细胞中β-catenin含量减少、β-catenin显示的荧光强度降低、TCF/LEF活性减弱。ITGB4BP还能显著抑制LiCl对β-catenin的激活作用。结论 ITGB4BP可以抑制Wnt/β-catenin信号通路的活性。 关键词：ITGB4BP；Wnt/β-catenin信号通路；LiCl ITGB4BP downregulates the activity of Wnt/β-catenin signaling pathway LAI Qing-wu1，2, PENG Xu2, TAN Jiang-lin2, ZHANG Xiao-rong2, HU Xiao-hong2, ZHANG Yi1, LUO Gao-xing2, WU Jun2 ( 1South China University of Technology, School of Bioscience and Bioengineering, Guangzhou 510006；2Institute of Burn Research, Chongqing Key laboratory of Proteomics of Diseases, State Key Laboratory of Trauma, Burns and Combined Injury, Southwest Hospital, Third Military Medical University, Chongqing 400038 ) Abstract: Objective To investigate the effects of ITGB4BP[integrin beta 4 binding protein] on the Wnt/β-catenin signaling pathway. Methods Human embryonic kidney cell line HEK293 was transfected with blank vector plasmid pCMV-3B as the control group and plasmid pCMV-ITGB4BP as the experiment group, respectively. β-catenin expression was checked by Western blot and laser confocal microscopy. Then the two groups with or without LiCl were cotransfected with plasmid TOPFlash containing promoter TCF/LEF and plasmid pRL-TK as the internal control, the TCF/LEF activity was analyzed by luciferase activity assay. Results As compared with the control group, β-catenin level was decreased significantly in HEK293 cells transfected with pCMV-ITGB4BP. The fluorescence intensity of β-catenin was weakened obviously and the TCF/LEF activity was reduced significantly. Moreover, The activation of LiCl on β-cate