EHBP1 Knockout Huh-7 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population derived from the human hepatocellular carcinoma cell line Huh-7, engineered for targeted disruption of the EHBP1 gene. This knockout model eliminates functional EHBP1 protein expression, enabling investigation of EHBP1-dependent mechanisms in a hepatic cancer background. The polyclonal format provides a heterogeneous knockout population suitable for pooled studies, avoiding clonal bias while maintaining target gene ablation. This loss-of-function system is designed for rigorous analysis of endocytic trafficking, actin dynamics, and insulin signaling in liver cancer cells.
The Huh-7 cell line, established in 1982 from a well-differentiated hepatocellular carcinoma of a Japanese male, is a widely used model for hepatocyte biology, hepatitis C virus replication, and anticancer drug screening. These epithelial cells retain key hepatic features and are permissive for HCV infection, making them invaluable for studying liver disease pathogenesis. Their robust proliferation and well-characterized signaling networks, including growth factor and metabolic pathways, provide a suitable platform for examining the role of endocytic adaptor proteins in cancer progression and metabolic regulation.
EHBP1 (EH domain-binding protein 1) functions as a scaffold linking clathrin-mediated endocytosis to the actin cytoskeleton. It interacts with EH domain-containing proteins such as Eps15 and Eps15R, as well as actin, thereby coupling vesicle formation to cytoskeletal dynamics. EHBP1 functions downstream of receptor tyrosine kinases like the EGF receptor and PDGF receptor, and it operates within a complex including Grb2, Cbl, and Dynamin-2. It also participates in insulin-stimulated GLUT4 translocation via PI3K/Akt signaling, regulating glucose uptake. Through its binding to Intersectin and Cortactin, EHBP1 promotes Arp2/3-mediated actin polymerization, influencing vesicle trafficking and cell migration.
In the Huh-7 hepatocellular carcinoma context, EHBP1 knockout is particularly relevant for dissecting the interplay between endocytosis, actin remodeling, and tumor cell behavior. Liver cancer cells rely on aberrant endocytic trafficking for growth factor receptor signaling, migration, and invasion. Disruption of EHBP1 may impair these processes, providing a tool to study mechanisms of metastasis and drug resistance. Additionally, given the liver’s central role in metabolism, this model allows examination of insulin signaling crosstalk in a hepatic cancer background, offering insights into cancer-associated metabolic alterations.
This polyclonal knockout cell population is suited for diverse experimental applications, including Western blotting and immunofluorescence to assess endocytic markers and actin organization, transferrin uptake assays to quantify receptor-mediated endocytosis, and insulin-stimulated GLUT4 translocation assays. Researchers can perform cell migration and invasion assays to evaluate metastatic potential, as well as RNA sequencing to identify transcriptomic changes resulting from EHBP1 loss. Co-immunoprecipitation studies can validate protein interaction networks involving Eps15, Dynamin-2, or Cortactin. For further information, please contact Ascent Research.
