The development of a functional assay system of Wnt/?-catenin signal pathway
- VernacularTitle:Wnt/?-连环蛋白信号途径活性的分析系统的建立
- Author:
Tao FENG
- Publication Type:Journal Article
- Keywords:
Wnt/?-catenin signaling pathway;
?-catenin and GFP fusion protein;
Functional assay system
- From:
Journal of Chongqing Medical University
1986;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To develop a Wnt-dependent ?-catenin-mediated functional assay system.Methods:To construct a vector,pCMV-bcGFP,which can express the ?-catenin and green fluorescent protein(GFP) fusion protein;transfect HEK293 with the vector,select the cells with G418 and LiCl and establish the stable cell line,293-bc-GFP by cloning.Results:293-bc-GFP stable cell line was developed;the background expression of ?-catenin-GFP fusion protein was detectable at a very low level in the cell line under fluorescent microscope and via Western blotting analysis;however,the fusion protein was showed to significantly increase in the presence of Wnt1 and lithium.Conclusion:Our results showed that the system is regulated by ?-catenin in a Wnt-responsive fasion.Thus,it can be used as a functional reporter to identify potential upstream factors during tumorigenesis,as well as to screen for the potential anti-cancer agents that specifically inhibit ?-catenin signaling in human tumors.
