Optimizations of a novel fluorescence polarization-based high-throughput screening assay for β-catenin/LEF1 interaction inhibitors.

Aberrant activation of the Wnt/β-catenin signaling pathway is prominent in the development and metastasis of non-small cell lung cancer (NSCLC). Highly effective inhibition of this pathway highlights a therapeutic avenue against NSCLC. Moreover, β-catenin/LEF1 interaction regulates β-catenin nuclear transport as well as the transcriptions of the key oncogenes in Wnt/β-catenin signaling pathway. Therefore, interruption of this interaction would be a promising therapeutic strategy for NSCLC metastasis. To date, no economical and rapid high-throughput screening (HTS) assay has been reported for the discovery of β-catenin/LEF1 interaction inhibitors. In this study, we developed a novel fluorescence polarization (FP)-based HTS assay to identify β-catenin/LEF1 interaction inhibitors. The FITC-LEF1 sequence, incubation time, temperature, and DMSO resistance were optimized, and then a high Z' factor of 0.77 was achieved. A pilot screening of a natural product library via this established FP screening assay identified sanguinarine analogues as potential β-catenin/LEF1 interaction inhibitors. GST pull-down and surface plasmon resonance (SPR) assay demonstrated that β-catenin/LEF1 interaction is a potential anticancer target of sanguinarine in vitro. This newly developed FP screening assay will be vital for the rapid discovery of novel Wnt inhibitors targeting β-catenin/LEF1 interaction.

Chen Y ，Fu Z ，Li D ，Yue Y ，Liu X ... -  《-》

[Optimizations of an ELISA-like high-throughput screening assay for the discovery of β-catenin/TCF4 interaction antagonists].

Fu Z ，Yan G ，Zhu X ，Liu X ，Chen Y ... -  《-》

[Development of an ELISA-based high throughput screening method for novel anticancer agents targeting β-catenin/Lef1 interaction].

Chen Y ，Niu X ，Li Y ，Liu X ... -  《-》

Analysis of the interaction of BCL9 with beta-catenin and development of fluorescence polarization and surface plasmon resonance binding assays for this interaction.

Kawamoto SA ，Thompson AD ，Coleska A ，Nikolovska-Coleska Z ，Yi H ，Wang S ... -  《-》

Development of a novel fluorescence polarization-based assay for studying the β-catenin/Tcf4 interaction.

Tian W ，Xu Y ，Han X ，Duggineni S ，Han X ，Huang Z ，An J ... -  《-》