The EHD4 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the SK-HEP-1 human hepatocarcinoma line, featuring targeted disruption of the EHD4 gene. This heterogeneous pool provides a robust loss-of-function tool for studying EHD4-mediated processes without clonal artifacts.
SK-HEP-1 is a human liver adenocarcinoma cell line with hepatocyte-like characteristics, widely used in hepatocellular carcinoma research and metabolic studies. Its epithelial morphology and hepatic gene expression profile make it a relevant model for tumor biology and diabetes-related investigations.
EHD4 encodes an ATPase critical for endocytic recycling. By utilizing ATP hydrolysis, EHD4 oligomerizes on recycling endosomes to drive membrane tubulation and vesicle formation, enabling the return of internalized receptors to the plasma membrane. It operates downstream of insulin and growth factor signaling and is regulated by Rab GTPases. EHD4 directly interacts with Rab11, Rab8, EHBP1, and the Arp2/3 complex, linking receptor recycling to the actin cytoskeleton. These interactions control surface levels of transferrin receptor and integrins, as well as GLUT4 translocation, thereby modulating cell migration, invasion, and glucose uptake. EHD4 collaborates with family members EHD1, EHD2, and EHD3 to maintain endocytic traffic.
Given EHD4’s role in integrin recycling and glucose transporter translocation, its knockout in SK-HEP-1 cells provides a powerful model to dissect the molecular drivers of hepatocarcinogenesis and type 2 diabetes-associated metabolic dysregulation. Reduced surface integrin expression may impair focal adhesion dynamics and inhibit cell motility, while defective GLUT4 trafficking could diminish glucose uptake, mirroring insulin resistance phenotypes.
For instance, flow cytometry can quantify surface levels of transferrin receptor and integrins, while transwell assays assess migratory and invasive potential. Immunofluorescence analysis can reveal EHD4-dependent alterations in endosomal distribution and actin cytoskeleton organization. Glucose uptake measurements using labeled 2-deoxyglucose can monitor GLUT4 trafficking defects. Co-immunoprecipitation experiments can confirm loss of interaction with Rab11, Rab8, or EHBP1, and global expression profiling can uncover compensatory changes. These polyclonal cells also serve as an effective platform for functional genomic screens and pharmacological modulation of endocytic recycling pathways. For additional technical information, please contact Ascent Research.
