The CCDC25 Knockout NCI-H1975 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population for studying CCDC25 function in a human lung adenocarcinoma background. This knockout model was generated by targeting the CCDC25 gene in the NCI-H1975 host cell line, creating a pool of cells with heterogeneous gene disruption. The polyclonal format enables investigation of CCDC25-dependent phenotypes without the clonal selection artifacts that can arise in single-cell-derived knockouts, making it particularly suitable for metastasis and cell migration research where population-level behavior is critical. This product is supplied as a ready-to-use cryopreserved pool, supporting straightforward culture and assay implementation.
NCI-H1975 is a widely characterized human non-small cell lung adenocarcinoma epithelial cell line derived from a non-smoker female. It harbors endogenous EGFR L858R and T790M mutations, representing a clinically relevant model for EGFR-targeted therapy resistance. The cells exhibit adherent growth, forming epithelial-like monolayers with typical lung adenocarcinoma features. The L858R mutation confers increased EGFR tyrosine kinase activity, while T790M reduces inhibitor binding, mimicking drug resistance seen in patients. This genetic background is essential for examining the interplay between oncogenic EGFR signaling and CCDC25-mediated metastatic processes in lung adenocarcinoma.
CCDC25 encodes a transmembrane receptor that specifically recognizes neutrophil extracellular trap DNA (NET-DNA) in the tumor microenvironment. Upon ligand binding, CCDC25 activates integrin-linked kinase (ILK) signaling, leading to ??-parvin recruitment, focal adhesion kinase phosphorylation, and actin cytoskeleton reorganization. Representative pathway components include NET-DNA, CCDC25, ILK, ??-parvin, paxillin, and cortactin. The receptor is activated by extracellular NET-DNA generated during NETosis, which can be stimulated by inflammatory agents such as PMA or LPS. Downstream, CCDC25 promotes cell migration and metastasis by driving ILK autophosphorylation and subsequent ??-parvin-mediated cytoskeletal remodeling. This signaling axis integrates external NET signals with intracellular migratory machinery.
In the NCI-H1975 background, CCDC25 knockout provides a powerful tool to dissect the contribution of NET-driven signaling to lung adenocarcinoma metastasis. The EGFR T790M/L858R mutant model recapitulates advanced NSCLC with acquired EGFR inhibitor resistance, a clinical scenario often associated with increased metastasis. By ablating CCDC25 function, researchers can investigate how NETosis in the tumor microenvironment influences cancer cell motility, invasion, and metastatic seeding independently of EGFR signaling. This knockout model is especially relevant for exploring the link between inflammatory NET responses and EGFR-mutant lung cancer progression, and for evaluating CCDC25 as a potential therapeutic target.
This polyclonal knockout cell population is ideally suited for a range of advanced research applications, including Transwell migration and invasion assays to quantify CCDC25-dependent motility, NETosis induction experiments with PMA or LPS to assess NET-DNA-dependent signaling, and Western blot analysis of ILK and ??-parvin phosphorylation status. Co-immunoprecipitation can be employed to examine ILK interaction partners, while immunofluorescence allows visualization of NET structures and CCDC25 localization. Additional applications include phospho-signaling profiling to map pathway alterations and CRISPR-Cas9 knockout validation by genomic and protein analysis. These assays collectively enable rigorous dissection of the ILK-??-parvin signaling module downstream of CCDC25. For further information, contact Ascent Research.
