The ASPH Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human liver adenocarcinoma cell line, designed to disrupt the aspartate ??-hydroxylase (ASPH) gene. This polyclonal population offers a heterogeneous pool of cells with targeted gene disruption, enabling researchers to study loss-of-function effects without clonal selection artifacts.
SK-HEP-1 is a widely used hepatic adenocarcinoma cell line established from the ascites of a 52-year-old male with liver adenocarcinoma. These cells exhibit both epithelial and endothelial characteristics, making them a unique model for studying tumor cell plasticity, migration, and invasion in hepatocellular carcinoma. Their dual phenotype provides a relevant context for investigating ASPH-mediated signaling in liver cancer progression.
ASPH encodes aspartate ??-hydroxylase, a dioxygenase that hydroxylates aspartic acid residues in EGF-like domains of proteins such as Notch receptors. This modification enhances Notch receptor activation and downstream signaling. ASPH is regulated by EGFR signaling, Wnt signaling, and hypoxia, and requires Fe2+ and ??-ketoglutarate as cofactors. Hydroxylated Notch receptors interact with ligands DLL4 and JAG1, leading to ??-secretase cleavage and Notch intracellular domain release. The intracellular domain forms a complex with CSL to activate transcription of targets like Hes1, promoting cell motility and invasion. ASPH thus integrates upstream EGFR and Wnt signals to potentiate Notch-dependent tumor progression, with crosstalk to AKT and MAPK pathways.
In SK-HEP-1 cells, ASPH is implicated in promoting the invasive and metastatic phenotype characteristic of liver adenocarcinoma. Disruption of ASPH in this polyclonal knockout population provides a valuable tool to dissect the mechanisms by which ASPH coordinates Notch and EGFR signaling to drive tumor aggressiveness. Given the endothelial-like features of SK-HEP-1, these cells also enable study of ASPH’s role in tumor-endothelial interactions. This model is relevant for hepatocellular carcinoma, cholangiocarcinoma, pancreatic cancer, and glioblastoma, where ASPH overexpression associates with poor prognosis.
Researchers can employ this polyclonal knockout model in functional assays, including western blotting and RT-qPCR to confirm ASPH disruption, migration and invasion assays to assess metastatic potential, and Notch reporter assays to quantify pathway activity. Phospho-signaling analyses can map changes in EGFR-AKT-MAPK activation. This cell population is suitable for anticancer drug screening targeting ASPH-dependent signaling and for functional genomics studies of EGFR-Wnt-Notch crosstalk. For further information or to discuss custom applications, please contact Ascent Research.
