The HCAR2 Knockout KYSE-30 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the KYSE-30 human esophageal squamous cell carcinoma cell line, designed to ablate expression of the HCAR2 gene. This polyclonal pool enables loss-of-function studies of the Gi/o-coupled receptor, which is physiologically activated by niacin and the ketone body beta-hydroxybutyrate. The absence of clonal selection preserves population-level heterogeneity, making the model suitable for dissecting HCAR2-dependent signaling pathways and cellular responses in a cancer-relevant context.
The parental KYSE-30 cell line was established from a well-differentiated invasive esophageal squamous cell carcinoma of a 64-year-old Japanese male. As a widely utilized epidermal epithelial tumor model, KYSE-30 recapitulates key features of esophageal cancer biology, including robust proliferative capacity and invasive potential. This background provides a relevant epithelial platform for investigating the role of HCAR2 in tumor cell-intrinsic signaling and the crosstalk between metabolic and inflammatory pathways.
HCAR2 is a G??i/o-coupled receptor activated by niacin and beta-hydroxybutyrate that suppresses adenylyl cyclase activity, reduces cAMP levels, and attenuates PKA signaling. Its engagement triggers G?¦? and beta-arrestin1/2-mediated pathways, including ERK1/2 phosphorylation and NF-??B inhibition, which downregulate pro-inflammatory cytokines such as TNF-alpha and IL-6. Key interacting partners include GRK2/5 for receptor desensitization and caveolin-1 for membrane compartmentalization. Downstream, HCAR2 modulates hormone-sensitive lipase and perilipin in lipolytic contexts, while in epithelial cells, it tunes transcriptional programs and cytokine secretion.
In the KYSE-30 esophageal squamous carcinoma model, HCAR2 signaling may intersect with oncogenic networks and tumor microenvironment interactions. Given that the receptor mediates anti-inflammatory and metabolic signals, its disruption in this polyclonal knockout population allows researchers to investigate how niacin or ketone body sensing influences cancer cell proliferation, survival, and migration. Because HCAR2 is expressed in various epithelial malignancies, this knockout tool provides a versatile system to study tumor-intrinsic functions of HCAR2 without the confounding effects of clonal variation.
The HCAR2 Knockout KYSE-30 Polyclonal Cells are suitable for a broad range of functional assays, including cAMP accumulation measurements to assess Gi-coupled signaling, western blotting and RT-qPCR for verification of knockout and downstream target expression, and cell-based assays such as MTT/BrdU proliferation, Transwell migration/invasion, and Annexin V apoptosis. Cytokine profiling via ELISA for TNF-alpha and IL-6 can delineate anti-inflammatory roles, while phospho-ERK analysis and RNA-seq enable deeper mechanistic dissection. For detailed protocols, technical support, or bulk order inquiries, please contact Ascent Research.
