The EHD4 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the EHD4 gene is disrupted in the AGS human gastric adenocarcinoma epithelial cell line. This loss-of-function model provides a heterogeneous pool of cells that avoids clonal selection biases, enabling robust assessment of EHD4-dependent phenotypes in a gastric epithelial context. The polyclonal format captures the collective impact of gene disruption on endocytic recycling and cell migration pathways.
The AGS cell line was established from a gastric adenocarcinoma of a 54-year-old female patient and is widely used in gastrointestinal research. It serves as a model for studying gastric cancer biology, mucosal barrier function, and Helicobacter pylori infection. Its epithelial origin and disease relevance make it particularly suitable for examining genes involved in tumor progression and metastasis.
EHD4 is an ATPase that catalyzes membrane tubulation and vesicle scission at recycling endosomes, facilitating the return of internalized receptors and adhesion molecules to the plasma membrane. Its activity is regulated by EGFR, phosphoinositides (e.g., PI3P), and small GTPases Rab4 and Rab11, and it interacts with cortactin, Arf6, Rab11-FIP2, and EHBP1 to coordinate actin dynamics. Downstream, EHD4 controls surface expression of ??1-integrin and the transferrin receptor, thereby governing cell adhesion, migration, and signal transduction. Dysregulation of EHD4 impairs polarized migration and promotes invasive behavior.
In gastric cancer, aberrant endocytic recycling contributes to invasion and metastasis. EHD4 loss in AGS cells provides a relevant model to dissect how disrupted integrin and receptor trafficking alters adhesion, migration, and signaling in gastric epithelial cells. This system can uncover mechanisms by which EHD4 deficiency may drive cancer cell dissemination, guiding therapeutic targeting strategies.
Researchers can employ these cells in Transwell migration/invasion assays, wound healing, and transferrin uptake experiments to assess endocytic trafficking and motility. Biochemical analyses via Western blotting, immunofluorescence, and co-immunoprecipitation allow examination of EHD4 complexes and targets like ??1-integrin. Proliferation (MTT) and apoptosis (Annexin V/PI) assays evaluate broader cellular consequences. For further details, please contact Ascent Research.
