The HCAR2 Knockout NCI-H1299 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of human NCI-H1299 non-small cell lung carcinoma cells harboring disruption of the HCAR2 gene. This heterogeneous pool of loss-of-function alleles lacks functional HCAR2 protein expression, providing a robust model system for studying HCAR2-dependent biology without clonal selection artifacts.
The parental NCI-H1299 cell line is an epithelial model of metastatic lung adenocarcinoma, originally isolated from a lymph node metastasis of a 43-year-old male patient. Widely used to study tumor cell migration, invasion, and metastatic colonization, NCI-H1299 cells offer a relevant background for examining how metabolic and immune signals intersect with lung cancer progression.
HCAR2 encodes a Gi/o-coupled receptor activated by nicotinic acid (niacin), the ketone body ??-hydroxybutyrate, and short-chain fatty acids such as butyrate. Ligand binding triggers G??i/o-mediated inhibition of adenylyl cyclase, lowering intracellular cAMP and PKA activity. This suppresses downstream effector pathways, notably NF-??B and ERK1/2, while also modulating Akt. HCAR2 signaling is further regulated by ??-arrestin-1/2 recruitment and GRK2-mediated receptor phosphorylation. Known physiological outcomes include anti-inflammatory macrophage polarization, adipocyte lipolysis inhibition, and vasodilation through prostaglandin D2 synthase and IL-10. In knockout cells, elimination of HCAR2 abrogates Gi-mediated cAMP suppression, resulting in de-repression of NF-??B and ERK signaling, which can amplify pro-inflammatory cytokine production and promote cell proliferation and motility.
In the context of NCI-H1299 metastatic lung cancer cells, HCAR2 knockout creates a unique model to investigate how metabolic signals such as niacin and butyrate influence tumor aggressiveness. Loss of the receptor??s anti-inflammatory restraint permits dissection of HCAR2-dependent contributions to invasive behavior, and enables testing of pharmacological agents that target this pathway in lung adenocarcinoma. Moreover, the model facilitates studies on metabolic-immune crosstalk relevant to hyperlipidemia and cancer progression.
Key experimental applications include Western blotting and quantitative RT-PCR to validate HCAR2 disruption and monitor NF-??B target gene expression, cAMP assays to quantify receptor function, phospho-ERK flow cytometry, and cytokine ELISA for IL-8 and IL-10. Functional assays such as wound healing and transwell invasion allow assessment of migration and invasion phenotypes. These polyclonal knockout cells are ideally suited for niacin and ??-hydroxybutyrate dose-response studies to discriminate receptor-mediated from off-target effects in lung cancer. For further information, please contact Ascent Research.
