DVL3 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human lung adenocarcinoma cell line NCI-H1975. In this pool, the DVL3 gene is disrupted through CRISPR/Cas9-mediated gene disruption, generating a heterogeneous loss-of-function model ideal for investigating Wnt signal transduction in lung cancer. These polyclonal cells enable interrogation of both canonical and non-canonical Wnt pathways in a population context, avoiding clonal selection bias.
The NCI-H1975 host cell line is a widely used non-small cell lung cancer (NSCLC) model, originally isolated from a female patient with lung adenocarcinoma. It harbors EGFR L858R and T790M mutations, which confer sensitivity to EGFR tyrosine kinase inhibitors and are linked to drug resistance. This background is valuable for studying crosstalk between EGFR and Wnt pathways and for testing combination therapies targeting both dependencies.
DVL3 is a cytoplasmic phosphoprotein and central Wnt signaling mediator, acting downstream of Frizzled (FZD) receptors and LRP5/6 co-receptors. Activated by Wnt ligands (e.g., Wnt3a, Wnt5a), DVL3 is phosphorylated by CK1?? and recruits AXIN1 and GSK3??, inhibiting the ??-catenin destruction complex, thereby stabilizing ??-catenin. Nuclear ??-catenin partners with TCF/LEF transcription factors to induce targets like MYC and CCND1. In non-canonical pathways, DVL3 signals through RHOA and RAC1 to control planar cell polarity and migration, and via JNK in the Wnt/Ca2+ pathway.
In NCI-H1975 cells, aberrant Wnt signaling contributes to tumor maintenance, EMT, and EGFR inhibitor resistance. Disrupting DVL3 attenuates ??-catenin/TCF-mediated transcription and non-canonical migration pathways, providing a system to dissect how DVL3-dependent Wnt signaling cooperates with mutant EGFR in NSCLC. This model helps assess how the loss of DVL3 alters sensitivity to EGFR inhibitors, Wnt inhibitors, or chemotherapeutics.
Applications include Western blotting for ??-catenin and phospho-LRP6, RT-qPCR for MYC and CCND1, and TOP/FOP reporter assays for ??-catenin/TCF activity. Functional assays encompass wound healing, transwell migration/invasion, immunofluorescence for DVL3 localization, and flow cytometry for cell cycle. Drug sensitivity profiling with Wnt inhibitors or EGFR TKIs can evaluate combination effects. For more information, contact Ascent Research.
