The ATP9A Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the ATP9A gene in the human hepatic adenocarcinoma cell line SK-HEP-1. This loss-of-function model enables the study of ATP9A-dependent phospholipid translocation and its roles in endosomal trafficking, cell migration, and cytokinesis within a hepatocellular carcinoma background. The polyclonal format provides a heterogeneous pool, reducing clonal adaptation artifacts.
SK-HEP-1 is an established hepatic adenocarcinoma cell line derived from ascitic fluid of a patient with hepatocellular carcinoma. It displays migratory and invasive traits typical of tumorigenic epithelial cells, making it a relevant model for metastasis and membrane trafficking studies. The cell line’s endosomal trafficking machinery is intact, allowing robust assessment of ATP9A-mediated lipid flippase activity.
ATP9A functions as a P4-ATPase flippase that actively transports aminophospholipids such as phosphatidylserine and phosphatidylethanolamine from the exoplasmic to the cytoplasmic leaflet of endosomal membranes. This activity is essential for maintaining membrane asymmetry, facilitating membrane curvature during vesicle formation, and promoting endosomal tubulation. It interacts obligately with CDC50A and CDC50B, which are required for its ER export and flippase activity. Upstream signals including cell cycle regulators, lipid signaling pathways, and growth factor stimulation modulate ATP9A activity. Downstream, ATP9A-mediated phospholipid redistribution influences actin cytoskeletal reorganization, supports contractile ring assembly during cytokinesis, and drives endosomal recycling of integrins and other cargo, thereby regulating cell migration and adhesion.
In the SK-HEP-1 hepatocellular carcinoma context, ATP9A disruption impairs endosomal recycling and reduces cell migration, potentially attenuating metastatic behavior. This knockout model enables dissection of how phospholipid asymmetry and membrane dynamics contribute to liver cancer cell motility. It represents a valuable tool for exploring the mechanistic role of ATP9A in tumor progression and the wider endosomal trafficking network.
These polyclonal knockout cells are ideal for cell migration assays such as wound healing or transwell migration, endocytosis and recycling assays using fluorescent ligands, immunofluorescence microscopy for endosomal markers, and molecular analyses including western blotting and RT-qPCR. They can be utilized in rescue experiments by re-expressing wild-type or mutant ATP9A or in chemical screens to identify regulators of phospholipid flippase activity. The polyclonal population format provides a robust and reproducible model that mitigates single-cell clonal artifacts. For further technical details or purchase inquiries, please contact Ascent Research.
