The EHD3 Knockout NCI-H1975 Polyclonal Cells product comprises a CRISPR/Cas9-edited heterogeneous cell pool carrying targeted disruption of the EHD3 gene in the human NCI-H1975 lung adenocarcinoma cell line. This polyclonal knockout population preserves genetic diversity while abrogating EHD3 protein function, making it well-suited for studies that benefit from pooled cell responses or avoidance of clonal artifacts. Cells are provided as a characterization-verified, mycoplasma-negative, ready-to-use model for advanced endocytic trafficking and cancer research applications.
NCI-H1975 is a widely used non-small cell lung cancer (NSCLC) line derived from a lung adenocarcinoma patient. It harbors activating EGFR L858R and T790M mutations, conferring sensitivity to third-generation EGFR tyrosine kinase inhibitors (TKIs) while mediating resistance to first-generation inhibitors. This genetic background establishes NCI-H1975 as a clinically relevant model for studying oncogenic EGFR signaling, acquired TKI resistance, and tumor cell maintenance mechanisms.
EHD3 catalyzes membrane scission on recycling endosomes, facilitating the return of integrins and EGFR to the cell surface. It functions downstream of Rab4 and Rab11, and forms complexes with EHD1/2/4, actin, Arf6, and PACSIN2. Upstream activation by EGFR and PI3K/AKT signaling regulates EHD3 activity. Consequently, EHD3 influences integrin-mediated adhesion, cell motility, and EGFR signaling dynamics at the plasma membrane.
In NCI-H1975 cells, EHD3 loss is predicted to disrupt integrin recycling, impairing cell adhesion and directional migration??processes frequently dysregulated in metastatic NSCLC. Concurrently, altered EGFR recycling may affect signal duration and strength, potentially modifying TKI sensitivity and downstream PI3K/AKT and MAPK pathway activation. This polyclonal knockout model therefore provides a powerful system to dissect how endocytic recycling intersects with mutant EGFR signaling and metastatic behavior in a clinically relevant NSCLC background.
Research applications include fluorescence imaging of integrin and EGFR localization, flow cytometry for surface receptor quantification, and transwell migration/invasion assays. Co-immunoprecipitation studies can confirm EHD3 interactions with Rab4/Rab11 under various conditions. Phospho-EGFR western blotting and live-cell signaling analyses can probe how EHD3 loss alters activation kinetics, while high-content screens can identify endocytosis modulators selective for mutant EGFR cells. For further technical information, please contact Ascent Research.
