The ATP2B2 Knockout SK-HEP-1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human SK-HEP-1 hepatocellular carcinoma line. This heterogeneous pool contains cells with targeted disruption of the ATP2B2 gene, which encodes plasma membrane calcium-transporting ATPase 2 (PMCA2). The polyclonal format avoids clonal selection, providing a robust loss-of-function model for studying ATP2B2-dependent processes in a liver cancer background.
SK-HEP-1 cells were originally isolated from the ascites of a hepatic adenocarcinoma patient and are frequently employed as an epithelial model that shares features with liver sinusoidal endothelial cells. Their adherent growth and stable karyotype facilitate a wide range of in vitro assays, including high-resolution imaging and functional analyses, making them a versatile platform for investigating hepatic calcium biology and tumor pathophysiology.
ATP2B2 encodes PMCA2, a high-affinity calcium pump that actively extrudes Ca2+ from the cytoplasm, maintaining low resting calcium levels. PMCA2 activity is regulated by calmodulin binding and phosphorylation by CaMKII in response to upstream signals, including Wnt ligands. Downstream, PMCA2-mediated calcium clearance influences the activation of NFAT and CREB transcription factors and the protease calpain. The pump also interacts with scaffold proteins such as NHERF and PDZ domain-containing proteins, linking calcium efflux to localized signaling. Disruption of ATP2B2 therefore perturbs the Wnt/Frizzled, PLC/IP3, and CaMKII/NFAT pathways, altering gene expression programs.
In hepatocellular carcinoma, dysregulated calcium signaling drives proliferation, migration, and evasion of apoptosis. Loss of PMCA2 in SK-HEP-1 cells provides a cellular model to dissect how impaired calcium extrusion affects these processes, particularly through NFAT and CREB-dependent transcription. The model also holds relevance for calcium dysregulation disorders, as ATP2B2 mutations are associated with autosomal recessive deafness 12. Thus, this knockout system bridges calcium transport and oncogenic signaling in a disease-relevant context.
These polyclonal knockout cells are suitable for calcium imaging with Fluo-4 to monitor intracellular dynamics, western blotting to confirm PMCA2 protein ablation, and functional assays such as cell viability, migration, and invasion tests. Flow cytometry enables quantification of apoptosis. Typical research applications include mechanistic studies of calcium signaling in hepatocellular carcinoma, investigation of PMCA2 as a therapeutic target, and screening of calcium-modulating compounds. For further details, please contact Ascent Research.
