The EHD4 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human non-small cell lung carcinoma (NSCLC) cell line NCI-H1299, featuring a targeted disruption of the EHD4 gene. This gene-edited product provides a versatile loss-of-function model for investigating the role of EHD4 in endocytic recycling, receptor trafficking, and cellular processes associated with cancer metastasis. As polyclonal knockout cells, the population harbors a heterogeneous mix of CRISPR-mediated gene disruptions, enabling robust study of EHD4 function without clonal selection artifacts.
The parental NCI-H1299 cell line was originally established from a lymph node metastasis of a lung adenocarcinoma patient, making it a widely used model for studies on NSCLC metastasis, tumor cell migration, and invasion. NCI-H1299 cells lack functional p53 and exhibit high tumorigenic and metastatic potential, rendering them particularly suitable for assessing the contribution of EHD4 to malignant phenotypes. Their origin from a metastatic site provides a physiologically relevant context for examining endocytic trafficking pathways that drive cancer dissemination.
EHD4 encodes an EH domain-containing protein that functions as a key regulator of endocytic recycling, controlling the return of internalized membrane receptors to the cell surface. It interacts with Rab11 on early endosomes to sort cargo including EGFR and ??1 integrin into recycling tubules. Upstream, EGF stimulation and integrin activation modulate EHD4, with Src kinase phosphorylation regulating its membrane association. Downstream, EHD4-mediated recycling sustains EGFR signaling and delivers integrins to the surface, promoting migration and invasion. Additionally, EHD4 coordinates with the Arp2/3 complex to remodel the actin cytoskeleton during trafficking. Disruption of EHD4 impairs receptor recycling, attenuating signaling and reducing migratory capacity.
In the NCI-H1299 background, knockout of EHD4 disrupts critical trafficking pathways that sustain the metastatic behavior of lung adenocarcinoma cells. By impairing the recycling of EGFR and ??1 integrin, the model compromises key signaling axes that promote tumor cell migration and invasion, thereby providing a tool to dissect the molecular mechanisms underlying NSCLC metastasis. This polyclonal knockout population is particularly valuable for comparing the pooled effects of various genetic disruptions and mitigating single-clone biases, reflecting the heterogeneous nature of tumors. The model enables researchers to examine how loss of EHD4 function modulates responses to growth factor stimulation and adhesion-dependent signals within a metastatic NSCLC context.
This EHD4 knockout model is suited for studying endocytic recycling in cancer, investigating EGFR and integrin trafficking, and screening compounds targeting metastasis. Typical assays include western blotting for EHD4 loss, immunofluorescence for receptor localization, transwell migration and invasion assays, and flow cytometry for surface integrin levels. The cells can also be used to evaluate anti-metastatic drugs by monitoring changes in cell motility. For additional information on this product, please contact Ascent Research.
