The EHD4 Knockout MES-OV Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the MES-OV human ovarian cancer cell line. This product serves as a loss-of-function model for EHD4 (EH domain-containing protein 4), a pivotal factor in endocytic membrane trafficking. The polyclonal nature ensures a diverse array of gene disruptions, capturing population-level effects without clonal selection biases. This model is optimized for functional studies of endosomal recycling and its impact on ovarian cancer cell behavior.
The host MES-OV cell line is a well-characterized model of mesenchymal subtype ovarian carcinoma, originally established from a patient tumor. Cells exhibit a spindle-like morphology, enhanced invasiveness, and robust metastatic potential, mirroring the aggressive clinical features of this subtype. MES-OV cells harbor key oncogenic mutations and signal through MAPK/ERK and PI3K/AKT pathways, making them particularly suited for investigating the molecular underpinnings of cancer dissemination.
EHD4 orchestrates the recycling of internalized cell surface receptors, including integrins and receptor tyrosine kinases, back to the plasma membrane. It functions within a macromolecular complex involving ARF6, RAB11, RAB35, actin, and the closely related EHD1. By driving membrane tubulation and fission, EHD4 facilitates the return of receptors that have been activated by ligands such as EGF, TGF-??, and extracellular matrix components. This process sustains downstream signaling through the MAPK/ERK and PI3K/AKT cascades, ultimately promoting RAC1-dependent actin cytoskeleton remodeling. Disruption of EHD4 therefore leads to attenuated receptor trafficking and diminished output from these pro-migratory pathways.
In the mesenchymal MES-OV background, EHD4 knockout is expected to significantly impair the recycling-dependent signaling that fuels the cells’ invasive phenotype. By reducing the surface availability of key receptors, the knockout likely blunts the sustained activation of ERK and AKT, thereby compromising cell migration and invasion. This model thus provides a physiologically relevant platform to dissect the cross-talk between endocytic trafficking and oncogenic signaling in a highly aggressive ovarian cancer setting. It also enables exploration of how subcellular trafficking abnormalities contribute to metastasis and therapy resistance.
Typical applications for these polyclonal knockout cells encompass transwell migration and invasion assays, endocytosis and recycling measurements using fluorescent ligands, and phospho-signaling analyses by western blotting. Co-immunoprecipitation studies can map altered protein interactions, while immunofluorescence microscopy permits visualization of receptor distribution and actin architecture. Flow cytometry enables quantification of surface receptor abundance. Collectively, these assays support research into ovarian cancer metastasis, endocytic trafficking mechanisms, and anti-metastatic drug development. For more information, contact Ascent Research.
