The EHD2 Knockout NCI-H1975 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human NCI-H1975 lung adenocarcinoma line. This product introduces targeted disruption of the EHD2 gene through CRISPR/Cas9-mediated gene editing, resulting in a heterogeneous pool of cells with loss-of-function mutations. As a polyclonal population, it maintains genetic diversity and avoids clonal selection biases, making it suitable for pooled screening and bulk functional assays.
The NCI-H1975 host cell line is a widely used model of non-small cell lung cancer (NSCLC) derived from a 62-year-old female patient. It harbors activating mutations in the epidermal growth factor receptor (EGFR) gene??specifically L858R in exon 21 and T790M in exon 20??which confer oncogenic signaling and acquired resistance to first-generation tyrosine kinase inhibitors (TKIs). These cells exhibit epithelial morphology and are frequently employed to study EGFR-driven tumorigenesis, drug resistance mechanisms, and metastatic progression in lung adenocarcinoma.
EHD2 (Eps15 homology domain-containing protein 2) functions as a key regulator of clathrin-independent endocytosis and integrin trafficking. It interacts with caveolin-1, Arf6, syndapin2, and Eps15 to mediate recycling of integrin ??5??1 to the plasma membrane, essential for focal adhesion turnover and actin cytoskeleton reorganization. EHD2 acts downstream of EGFR and TGF-?? signaling and upstream of Rac1 activation and RhoA suppression, thus modulating actin polymerization and cell migration. Hypoxia also regulates EHD2 expression. Loss of EHD2 disrupts integrin recycling, impairing focal adhesion disassembly and cytoskeletal dynamics.
In NCI-H1975 cells with EGFR L858R/T790M mutations, EHD2 knockout allows dissection of endocytic trafficking in EGFR-driven metastasis. Persistent pro-migratory signaling from mutant EGFR may depend on EHD2-mediated integrin recycling for tumor invasion and TKI resistance. Disruption of EHD2 attenuates metastatic signaling by reducing surface integrin levels and impairing Rac1-dependent actin remodeling, thereby altering cell motility in this aggressive cancer model.
This product is suitable for investigating EHD2??s role in lung adenocarcinoma metastasis, EGFR TKI resistance, and endocytic trafficking in cancer. Typical assays include Western blotting and RT-qPCR for knockout validation, cell migration and invasion assays, immunofluorescence for integrin ??5??1 and focal adhesion markers, flow cytometry for surface integrin, and phospho-EGFR analysis. Co-immunoprecipitation can probe altered EHD2 interaction networks. For further information or to request a quote, please contact Ascent Research.
