ECE1 Knockout SK-HEP-1 Polyclonal Cells are a population of CRISPR/Cas9-edited human liver sinusoidal endothelial cells engineered for functional disruption of the ECE1 gene. This polyclonal knockout pool provides a heterogeneous loss-of-function model that eliminates the enzymatic conversion of inactive precursors into active vasoactive peptides, enabling robust interrogation of endothelin signaling without the need for single-cell clonal isolation.
SK-HEP-1 cells are derived from ascitic fluid of a male patient with liver adenocarcinoma and exhibit an endothelial phenotype, making them suitable for studies on liver sinusoidal endothelium, hepatic barrier function, angiogenesis, and metastasis. They retain key endothelial characteristics and are widely used as a model for liver endothelial biology and cancer-related vascular processes.
ECE1 encodes endothelin-converting enzyme-1, a transmembrane metalloprotease that processes big endothelin-1 (and -2, -3) to generate active endothelin-1, a potent vasoconstrictor, as well as cleaves other vasoactive peptides such as bradykinin and substance P. In the endothelin signaling pathway, ECE1-produced endothelin-1 binds to G protein-coupled receptors EDNRA and EDNRB, activating Gq/11 and downstream effectors including phospholipase C (PLC), inositol trisphosphate (IP3), diacylglycerol (DAG), calcium mobilization, and protein kinase C (PKC). This cascade triggers the MAPK and PI3K/AKT pathways, RhoA/ROCK signaling, and transcription factors such as AP-1, thereby regulating vasoconstriction, angiogenesis, cell proliferation, and vascular remodeling. ECE1 expression is modulated by upstream regulators including GATA2, HIF1A, TNF-alpha, TGF-beta, and hypoxia, while its activity converges on EDN1/EDNRA/EDNRB-mediated signaling, eNOS modulation, and cGMP-PKG pathways.
Disruption of ECE1 in SK-HEP-1 cells eliminates the generation of mature endothelin-1, leading to abrogation of endothelin receptor activation and downstream signaling events. This results in impaired vasoconstrictive responses, reduced angiogenic capacity, and blunted proliferative signals, thereby creating a defined genetic model to dissect endothelin-driven processes within the liver sinusoidal endothelial environment. The knockout enables direct assessment of ECE1-mediated effects on endothelial barrier integrity, angiogenesis, and the response to vasoactive mediators in a hepatic context.
Researchers can employ this polyclonal knockout model in assays such as endothelin-1 ELISA, RT-qPCR for EDN1/EDNRA/EDNRB, Western blotting of phospho-MAPK and phospho-AKT, tube formation and migration assays, calcium imaging, and TEER measurements for barrier function. It serves as a valuable tool for investigating the role of endothelin-1 in portal hypertension, liver fibrosis, and cancer metastasis, as well as for screening ECE1 inhibitors or evaluating therapeutic interventions like bosentan or ambrisentan. This product is ideal for mechanistic studies of vasoactive peptide metabolism and vascular pathology in the liver endothelium. For additional information, please contact Ascent Research.
