The ITPKC Knockout SK-HEP-1 Polyclonal Cells product provides a ready-to-use CRISPR/Cas9-edited polyclonal knockout cell population in the SK-HEP-1 background, with targeted disruption of the ITPKC gene. This polyclonal format captures the genetic heterogeneity inherent to CRISPR/Cas9-mediated gene disruption, enabling robust loss-of-function studies without the need for single-cell cloning. The cells are supplied as a pooled population directly amenable to functional assays, serving as a versatile in vitro model for investigating ITPKC-dependent signaling in hepatic and endothelial contexts.
SK-HEP-1 is a human liver adenocarcinoma cell line originally derived from the ascites of a patient with hepatocellular carcinoma. It displays a unique endothelial-like phenotype, characterized by expression of endothelial markers such as CD34 and von Willebrand factor, alongside epithelial features. This dual identity makes SK-HEP-1 a widely accepted model for liver sinusoidal endothelial cells while retaining hepatic carcinoma traits, offering a platform to study both endothelial biology and hepatocellular carcinoma pathophysiology in a single cell background.
ITPKC encodes inositol 1,4,5-trisphosphate 3-kinase C, which phosphorylates the second messenger IP3 to IP4, thereby reducing IP3-induced calcium release from the endoplasmic reticulum and dampening downstream calcium-dependent processes. The kinase is activated downstream of T cell receptor/CD3 stimulation and CD28 co-stimulation, as well as insulin and growth factor signals via PKC. ITPKC interacts with calmodulin and F-actin, and its activity modulates the localization and function of signaling complexes involving Rac1 and the IP3 receptor. By limiting cytosolic calcium transients, ITPKC negatively regulates calcineurin-mediated dephosphorylation and nuclear translocation of NFAT, a key transcription factor in immune and metabolic responses. In hepatic cells, ITPKC also intersects with insulin signaling: it influences AKT phosphorylation and downstream targets such as GSK3 and FOXO, thereby impacting gluconeogenic gene expression and glucose homeostasis.
Loss of ITPKC function in the SK-HEP-1 context disrupts the negative regulation of calcium flux and NFAT activation, while also perturbing AKT/FOXO-mediated insulin signaling. This combined disruption renders the knockout cells a powerful tool for dissecting the crosstalk between calcium signaling and metabolic pathways in a liver-derived endothelial-like cell model. Given SK-HEP-1??s expression of endothelial markers and hepatic lineage, the ITPKC knockout can illuminate mechanistic links between ITPKC-dependent calcium modulation, endothelial function, and hepatocellular carcinoma biology, including potential roles in tumor cell proliferation and insulin resistance.
These polyclonal knockout cells are suited for a broad range of research applications, including the study of Kawasaki disease pathophysiology, T-cell activation and calcium signaling, hepatic insulin resistance, and liver cancer metabolism. Experimental approaches such as calcium flux assays, western blotting for phospho-AKT and NFAT, RT-qPCR for cytokine expression, glucose production assays, and immunocytochemistry for NFAT localization are directly supported. The model can also be employed to explore endothelial cell biology and to screen for modulators of ITPKC-related signaling. For more information or to discuss custom applications, please contact Ascent Research.
