The EHD3 Knockout SK-HEP-1 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the expression of the EHD3 gene in the human SK-HEP-1 cell line. This gene-edited pool enables loss-of-function studies of EHD3 in a heterogeneous cell population, offering a robust model for investigating the functional consequences of EHD3 ablation without clonal selection.
The host SK-HEP-1 cell line is derived from the ascites of a patient with liver adenocarcinoma and serves as a well-established model for hepatocellular carcinoma (HCC), metastasis, and anticancer drug screening. These adherent epithelial cells retain characteristics of malignant liver-derived cells and are frequently employed to examine tumor cell motility, invasion, and metastatic dissemination in vitro.
EHD3 encodes an EH domain-containing ATPase that functions as a key regulator of endocytic recycling, facilitating the return of internalized receptors and adhesion molecules from endosomes back to the plasma membrane. EHD3 interacts with endocytic regulatory proteins including Rab5, Rab11, and Eps15, as well as the actin cytoskeleton via F-actin and syndapin. Through these interactions, EHD3 orchestrates the trafficking of integrins, transferrin receptors, and receptor tyrosine kinases, thereby modulating actin remodeling, cell adhesion, and downstream signaling pathways such as those mediated by Arf6.
In the context of hepatocellular carcinoma, disruption of EHD3 is expected to impair the recycling of adhesion and signaling receptors, potentially attenuating cell migration, invasion, and metastatic potential. The SK-HEP-1 polyclonal EHD3 knockout pool thus represents a valuable tool for dissecting the role of endocytic trafficking in liver cancer progression and for evaluating the dependency of HCC cells on EHD3-mediated membrane dynamics.
Researchers can utilize these polyclonal knockout cells in a variety of experimental settings. Endocytic recycling efficiency can be assessed via transferrin uptake and recycling assays, while changes in cell migration and invasion can be quantified using Boyden chamber or wound-healing assays. Surface levels of integrins and receptor tyrosine kinases can be measured by flow cytometry, and EHD3 disruption confirmed by western blotting and RT-qPCR. Immunofluorescence microscopy enables visualization of receptor mislocalization, and co-immunoprecipitation can be used to validate disrupted protein interactions. This product is particularly suited for high-throughput drug sensitivity screens and for studies exploring the intersection of vesicle trafficking and metastasis. For inquiries, contact Ascent Research.
