HCAR2 Knockout 786-O Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population of the human renal cell carcinoma line 786-O, engineered to ablate HCAR2 expression. The resulting loss-of-function model permits direct interrogation of niacin- and butyrate-responsive signaling in a cancer cell background without the genetic uniformity of a clonal line.
The parental 786-O line originates from clear cell renal cell carcinoma (ccRCC) and carries a mutant VHL gene, leading to constitutive HIF stabilization and pseudohypoxia. This widely used model recapitulates key features of kidney cancer, including altered metabolism and angiogenesis, and provides a susceptible background for studying nutrient-sensing receptors such as HCAR2.
HCAR2 encodes a G??i/o-coupled receptor activated by niacin, butyrate, and beta-hydroxybutyrate, leading to inhibition of adenylate cyclase and reduction of cAMP levels. Downstream, this modulates PKA, ERK1/2, AKT, NF-??B, and PPAR-?? activity, as well as adiponectin expression. The receptor interacts with Gi/o proteins (GNAI1, GNAI2, GNAO1) and is regulated by GRK2 and ??-arrestins. Consequently, HCAR2 mediates anti-lipolytic and anti-inflammatory responses, suppressing NF-??B and promoting PPAR-?? while influencing ERK and AKT signaling to affect cell survival and metabolism.
In 786-O cells, HCAR2 knockout allows dissection of GPCR signaling in cancer metabolism. The VHL-deficient ccRCC background, with its metabolic alterations, provides a relevant setting to examine how HCAR2 links extracellular nutrients like niacin and butyrate to energy sensing and inflammation. Loss of receptor function can uncover contributions to proliferation, metabolic reprogramming, and tumor microenvironment interactions under dietary or microbial metabolite conditions.
Applications include cAMP assays for Gi signaling validation, niacin/butyrate dose?Cresponse studies for metabolic and anti-inflammatory endpoints, and western blotting for phospho-ERK. RT-qPCR can assess transcriptional changes, while flow cytometry and migration/invasion assays explore immune and metastatic phenotypes. This model supports drug target validation for dyslipidemia, obesity, diabetes, and inflammatory bowel disease, as well as cancer metabolism and microbiota-host interaction studies. For ordering and support, contact Ascent Research.
