The HCAR2 Knockout SK-HEP-1 Polyclonal Cells product provides a population of CRISPR/Cas9-edited cells with targeted disruption of the HCAR2 gene. These polyclonal knockout cells are generated from the SK-HEP-1 liver adenocarcinoma cell line and offer a loss-of-function model without isolation of single clones. The edited population enables robust studies of HCAR2 function by abolishing receptor expression in a heterogeneous, yet well-characterized, host cell background. This polyclonal format preserves genetic diversity inherent to the parental line while ensuring efficient gene ablation for downstream assays.
SK-HEP-1 is a human liver adenocarcinoma cell line originally derived from the ascitic fluid of a male patient; it displays both epithelial and endothelial characteristics, making it a unique model for hepatocellular carcinoma (HCC) and liver vascular biology. The cell line exhibits features such as expression of endothelial markers alongside epithelial traits, allowing investigation of tumor cell plasticity and the tumor microenvironment. Its widespread use in HCC research includes studies on proliferation, migration, angiogenesis, and metabolic reprogramming, providing a relevant context for assessing HCAR2’s tumor-modulatory roles.
HCAR2, also known as GPR109A, is a Gi/o protein-coupled receptor activated by niacin, butyrate, and 3-hydroxybutyrate. Ligand binding triggers coupling to G??i/o proteins, inhibiting adenylyl cyclase to reduce cAMP. This leads to decreased PKA activity and inhibition of hormone-sensitive lipase, suppressing lipolysis. In immune and epithelial cells, HCAR2 activation attenuates NF-??B signaling, exerting anti-inflammatory effects. Downstream, the receptor engages ???arrestins and GRKs, and modulates ERK1/2 and Akt phosphorylation. Thus, HCAR2 integrates metabolic and inflammatory signals through a central Gi?cAMP?CPKA axis.
In the SK-HEP-1 liver adenocarcinoma model, HCAR2 expression may influence tumor cell behavior by linking lipid metabolism to inflammatory cascades. Butyrate, a product of gut microbial fermentation, and niacin, a dietary component, can signal through HCAR2 in the hepatic environment. Given SK-HEP-1’s endothelial-like phenotype, HCAR2 might also modulate vascular mimicry or interactions with the tumor stroma. Disrupting HCAR2 in these cells permits dissection of its contribution to HCC cell proliferation, survival, and migration under lipid?rich or inflammatory conditions, offering insights into the receptor’s potential as a therapeutic target in liver cancer and associated metabolic disorders.
This knockout model is ideally suited for a variety of experimental approaches. Researchers can perform Western blotting and cAMP assays to confirm HCAR2 ablation and altered second?messenger responses. Functional studies may include lipolysis assays, NF???B luciferase reporter assays, and cell proliferation or migration/invasion assays. Butyrate or niacin stimulation followed by phospho?signaling analysis (e.g., phospho?ERK1/2, phospho?Akt) and RNA?seq transcriptomics will elucidate HCAR2?dependent pathways in liver cancer. The model also supports drug?target validation for dyslipidemia, inflammatory bowel disease, and anti?tumor strategies. For additional information, please contact Ascent Research.
