The HEBP2 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the SK-HEP-1 human liver adenocarcinoma cell line, designed to disrupt HEBP2. This heterogeneous knockout model ablates HEBP2 protein across the cell pool, avoiding clonal bias while preserving phenotypic diversity. The loss-of-function mutations eliminate HEBP2??s heme-binding capacity, providing a robust tool for investigating heme metabolism, oxidative stress, and apoptosis in an endothelial-like hepatic context.
The SK-HEP-1 cell line, isolated from ascites of a liver adenocarcinoma patient, displays a mixed endothelial and epithelial/mesenchymal phenotype. It expresses liver sinusoidal endothelial markers, such as acetylated LDL uptake and tube formation, while retaining adenocarcinoma features. This immortalized line facilitates reproducible studies of endothelial biology, angiogenesis, and heme handling with liver-derived physiological relevance.
HEBP2 (SOUL) is a cytosolic heme-binding protein that sequesters free heme, preventing ROS generation via Fenton chemistry and suppressing apoptosis. Its primary interacting factor is heme, and it participates in a network with heme oxygenase 1 (HO-1) and cellular ROS homeostasis. Although upstream regulators and downstream targets remain unidentified, HEBP2 loss elevates free heme, heightening oxidative stress and apoptotic sensitivity, positioning it as a key cytoprotective factor in heme-rich environments.
In SK-HEP-1 cells, which encounter heme during hepatic endothelial function and pathological hemolysis, HEBP2 knockout markedly increases susceptibility to heme-induced cytotoxicity. The absence of HEBP2 buffering leads to elevated ROS and apoptosis upon heme challenge, modeling heme-mediated liver injury. This polyclonal knockout thus enables dissection of HEBP2??s protective role in endothelial-like cells and is relevant to hemolytic diseases, ischemia-reperfusion injury, and oxidative stress-linked liver pathologies.
Researchers can use these cells for quantitative ROS measurement (DCFDA assay), apoptosis detection (Annexin V/7-AAD), cell viability determination (MTT), and spectrophotometric heme-binding studies. Western blotting verifies HEBP2 ablation, and RT-qPCR analyzes oxidative stress-related transcripts. The polyclonal pool is ideal for drug screening targeting heme detoxification pathways and evaluating cytoprotective compounds. For further technical inquiries or ordering, please contact Ascent Research.
