Cell-based assay for low- and high-scale screening of the Wnt/β-catenin signaling modulators.

Deregulation of the Wnt/β-catenin signaling pathway is associated with many serious disorders, including cancer and Alzheimer's disease. The pivotal player is β-catenin, which avoids degradation after activation of the pathway and is translocated to the nucleus, where it interacts with TCF/LEF transcription factors and induces expression of genes involved in cell cycle and apoptosis regulation. The identification of small molecules that may affect Wnt/β-catenin signaling remains an important target during the development of novel therapies. We used the TCF/LEF lentiviral vector and the Wnt-independent H1703 cell line to develop a luciferase reporter-based cell assay for screening of the Wnt/β-catenin pathway modulators. Following the optimization of cell density, concentration of activator, and stimulation time, the reporter system was validated by demonstrating its specific and dose-dependent response to several established modulators of Wnt/β-catenin signaling such as Wnt3a, small interfering RNA (siRNA) against β-catenin, glycogen synthase kinase 3 (GSK-3), and β-catenin/TCF transcription complex inhibitors. Two pilot screens of inhibitors and activators of Wnt/β-catenin signaling identified potential novel modulators of this pathway. Our findings suggest that the H1703-7TFP assay constitutes a suitable model of low background and high sensitivity for the low- and high-scale screening of the Wnt/β-catenin pathway modulators.