Chen Minyong, Wang Jiangbo, Lu Jiuyi, Bond Michael C, Ren Xiu-Rong, Lyerly H Kim, Barak Larry S, Chen Wei
Department of Medicine, Duke University Medical Center, Durham, North Carolina 27710, USA.
Biochemistry. 2009 Nov 3;48(43):10267-74. doi: 10.1021/bi9009677.
Wnt proteins bind to seven-transmembrane Frizzled receptors to mediate the important developmental, morphogenetic, and stem cell related tissue-regenerative effects of Wnt signaling. Dysregulated Wnt signaling is associated with many cancers. Currently, there are no drug candidates or even tool compounds that modulate Wnt-mediated receptor trafficking, and subsequent Wnt signaling. We examined libraries of FDA-approved drugs for their utility as Frizzled internalization modulators, employing a primary imaged-based GFP fluorescence assay that uses Frizzled1 endocytosis as the readout. We now report that the anti-helminthic niclosamide, a drug used for the treatment of tapeworm, promotes Frizzled1 endocytosis, downregulates Dishevelled-2 protein, and inhibits Wnt3A-stimulated beta-catenin stabilization and LEF/TCF reporter activity. Additionally, following niclosamide-mediated internalization, the Frizzled1 receptor colocalizes in vesicles containing transferrin and agonist-activated beta(2)-adrenergic receptor. Therefore, niclosamide may serve as a negative modulator of Wnt/Frizzled1 signaling by depleting upstream signaling molecules (i.e., Frizzled and Dishevelled) and moreover may provide a valuable means of studying the physiological consequences of Wnt signaling.
Wnt蛋白与七次跨膜的卷曲受体结合,以介导Wnt信号通路在发育、形态发生以及与干细胞相关的组织再生方面的重要作用。Wnt信号通路失调与多种癌症相关。目前,尚无能够调节Wnt介导的受体转运及后续Wnt信号传导的候选药物甚至工具化合物。我们利用一种基于成像的主要绿色荧光蛋白(GFP)荧光检测方法(该方法以卷曲蛋白1的内吞作用作为读数),检测了美国食品药品监督管理局(FDA)批准的药物库,以评估其作为卷曲蛋白内化调节剂的效用。我们现在报告,抗蠕虫药物氯硝柳胺(一种用于治疗绦虫的药物)可促进卷曲蛋白1的内吞作用,下调散乱蛋白2的表达,并抑制Wnt3A刺激的β-连环蛋白稳定化以及淋巴样增强因子/转录因子(LEF/TCF)报告基因活性。此外,在氯硝柳胺介导的内化作用之后,卷曲蛋白1受体与含有转铁蛋白和激动剂激活的β2-肾上腺素能受体的囊泡共定位。因此,氯硝柳胺可能通过消耗上游信号分子(即卷曲蛋白和散乱蛋白)而作为Wnt/卷曲蛋白1信号通路的负调节剂,并且还可能为研究Wnt信号传导的生理后果提供一种有价值的手段。