The HCAR2 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the expression of the HCAR2 gene (also known as GPR109A) in the human CAL-27 cell line. This loss-of-function model enables researchers to investigate the biological roles of HCAR2 in a tongue squamous cell carcinoma background. The polyclonal knockout cells are generated using CRISPR/Cas9-mediated gene disruption, providing a heterogeneous population that can be used for various downstream assays to study HCAR2-dependent signaling pathways.
The parental CAL-27 cell line is derived from a human tongue squamous cell carcinoma and exhibits an adherent epithelial morphology. These cells are widely used as a model system for oral cancer research, including studies on tumor growth, invasion, and drug response. The epithelial origin of CAL-27 cells makes them particularly relevant for investigating the molecular mechanisms underlying head and neck squamous cell carcinoma, where HCAR2 signaling may play a modulatory role.
HCAR2 encodes a G protein-coupled receptor that is activated by niacin, butyrate, and other short-chain fatty acids such as beta-hydroxybutyrate. Upon ligand binding, HCAR2 couples to Gi/o proteins, leading to the inhibition of adenylyl cyclase and a subsequent decrease in intracellular cAMP levels. This signaling cascade influences several downstream pathways, including the MAPK/ERK pathway, NF-kB inhibition, and PPAR?? activation. Among the representative pathway components are the Gi alpha subunit, PKA, ERK1/2, and transcription factors like NF-kB and PPAR??. Additionally, receptor desensitization and internalization involve interacting factors such as GRKs and arrestins. Overall, HCAR2 mediates anti-lipolytic and anti-inflammatory responses, which are critical in metabolic regulation and immune modulation.
In the context of CAL-27 oral squamous carcinoma cells, HCAR2 knockout disrupts niacin- and butyrate-induced signaling, leading to altered cAMP dynamics and downstream effects on inflammatory and metabolic pathways. This disruption can impact cellular processes such as proliferation, migration, and inflammatory cytokine production. By ablating HCAR2 function, researchers can dissect its contribution to oral cancer progression and assess its potential as a therapeutic target. The model is particularly valuable for studying the intersection between metabolism and cancer, given the known roles of short-chain fatty acids in tumor microenvironments.
The HCAR2 Knockout CAL-27 Polyclonal Cells are suitable for a wide range of research applications, including GPCR signaling studies, cancer metabolism, and inflammatory response investigations. Commonly used assays with these cells include cAMP accumulation assays to measure receptor activity, western blotting for phosphorylated ERK1/2 to assess MAPK pathway activation, RT-qPCR for downstream target gene expression, and migration or invasion assays to evaluate metastatic potential. Additionally, these cells can be utilized in drug sensitivity screens to evaluate the efficacy of niacin, butyrate, or other HCAR2-targeting compounds. Immunofluorescence can further be employed to examine subcellular localization of signaling components. For additional information or to inquire about custom gene editing services, please contact Ascent Research.
