The EHD2 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the NCI-H1299 human lung carcinoma cell line, providing a loss-of-function model for studying EHD2 in caveolae-mediated endocytosis. The polyclonal pool allows investigation of EHD2 function without clonal artifacts and retains diverse genetic backgrounds, beneficial for robust statistical analysis. This model is suitable for a variety of functional assays to dissect EHD2-related signaling networks.
The NCI-H1299 cell line originates from a lymph node metastasis of a 43-year-old male with non-small cell lung carcinoma. These epithelial cells lack functional p53 and are widely used as a model for metastatic NSCLC, particularly for studies on cell migration, invasion, and anoikis resistance. Their aggressive phenotype and metastatic origin make them an ideal platform for exploring endocytic trafficking and cytoskeletal dynamics in lung cancer progression.
EHD2 is a dimeric ATPase that mediates caveolae-mediated endocytosis by binding membranes, inducing tubulation, and cooperating with dynamin2 for membrane scission. It stabilizes the caveolar coat through interactions with caveolin-1 and pacsin2/syndapin2, and links caveolae to the actin cytoskeleton via N-WASP and actin. EHD2 expression is regulated by MRTF-A/B and YAP/TAZ downstream of mechanical stretch and integrin signaling, and it governs actin polymerization, RhoA activity, focal adhesion turnover, and EGFR internalization. Disruption of EHD2 therefore dismantles caveolar organization and impairs downstream signaling and trafficking.
In NCI-H1299 cells, loss of EHD2 disrupts caveolae-dependent endocytosis, offering a system to scrutinize EGFR trafficking and its role in NSCLC metastasis. The knockout reveals how EHD2-mediated caveolar-cytoskeletal linkage affects cell stiffness, migration, and response to mechanical forces. This polyclonal population enables studies on anchorage-independent growth, transendothelial migration, and mechanotransduction, providing key insights into metastatic mechanisms. Moreover, the absence of clonal selection preserves the phenotypic heterogeneity typical of tumor populations, enhancing the translational relevance of experimental findings.
These cells are ideal for cholera toxin B uptake assays, Transwell and wound healing migration assays, and immunofluorescence analysis of caveolar and focal adhesion structures. They support phospho-EGFR signaling studies, proteomic identification of EHD2-dependent cargoes, and screening of caveolae-targeted therapeutics. The polyclonal EHD2 Knockout NCI-H1299 cells thus empower diverse experiments in cancer biology, endocytosis, and mechanobiology. For more information, please contact Ascent Research.
