HCAR2 Knockout HGC-27 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HGC-27 human gastric adenocarcinoma cell line, engineered to disrupt endogenous HCAR2 gene expression. This product provides a heterogeneous loss-of-function model, generated by introducing double-strand breaks and allowing non-homologous end joining to create diverse insertions or deletions across the polyclonal pool, without clonal selection. The resulting polyclonal knockout cells enable robust functional studies of HCAR2-dependent signaling in a gastric cancer background, avoiding single-clone artifacts. The product is supplied as a live proliferating culture, suitable for immediate experimental expansion and banking.
The HGC-27 host cell line is a poorly differentiated gastric adenocarcinoma line originally isolated from a metastatic lymph node of a patient with advanced gastric carcinoma. These cells exhibit epithelial morphology and are widely used to model gastric tumor biology, including metastatic potential, drug response, and inflammatory signaling. Their fast growth and stable genetic background make them an ideal platform for CRISPR-mediated gene disruption. The HCAR2 knockout in this context allows direct interrogation of metabolic-sensing receptor function in a clinically relevant tumor microenvironment.
HCAR2 (hydroxycarboxylic acid receptor 2, also known as GPR109A) is a metabolite-sensing G protein-coupled receptor activated by endogenous and dietary ligands, including nicotinic acid (niacin), the short-chain fatty acid butyrate, and the ketone body ??-hydroxybutyrate. Upon ligand binding, HCAR2 couples primarily to G??i/o proteins, leading to inhibition of adenylate cyclase, reduced intracellular cAMP levels, and consequent suppression of protein kinase A (PKA) activity. This signaling cascade attenuates the phosphorylation and activity of hormone-sensitive lipase (HSL), thereby inhibiting lipolysis in adipocytes. In immune and epithelial cells, reduced PKA activity leads to dampening of the NF-??B pathway, diminishing the expression of pro-inflammatory cytokines. Additionally, receptor activation promotes ??-arrestin recruitment, which can scaffold MAP kinase modules, contributing to context-dependent signaling outcomes.
In gastric adenocarcinoma, the role of HCAR2 is poorly defined, but emerging evidence suggests that gut microbiota-derived metabolites such as butyrate may influence tumor cell proliferation, apoptosis, and inflammation through GPCR-mediated mechanisms. The HGC-27 knockout model permits investigation of HCAR2-dependent effects on these processes, including potential tumor-suppressive or tumor-promoting actions. Given the receptor’s established anti-inflammatory roles in intestinal epithelia and immune cells, the model is also relevant for dissecting cross-talk between microbial metabolites and gastric tumourigenesis, with implications for dyslipidemia, inflammatory bowel disease, and cardiovascular disorders where HCAR2 is a therapeutic target.
Researchers can apply HCAR2 Knockout HGC-27 Polyclonal Cells to diverse experimental paradigms. Typical applications include comparative transcriptomic (RNA-seq) and proteomic profiling of wild-type versus knockout cells in the presence or absence of niacin, butyrate, or ??-hydroxybutyrate to map HCAR2-dependent pathways. Functional assays such as cAMP measurement, NF-??B reporter assays, cell viability, and migration/invasion assays permit phenotypic characterization. Co-immunoprecipitation can explore HCAR2 interactions with G??i, ??-arrestins, and downstream effectors. This model aids in validating HCAR2 as a node linking metabolism and inflammation in gastric cancer and in screening novel agonists for therapeutic intervention. For further information, please contact Ascent Research.
