The HCAR2 Knockout Ca Ski Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal cell pool with targeted disruption of the HCAR2 gene, designed for loss-of-function studies in a human cervical carcinoma model. The polyclonal format avoids clonal bias, offering a genetically diverse population that mimics the heterogeneity of tumor cell populations. This stable knockout model enables robust phenotypic analysis of HCAR2-dependent processes in a physiologically relevant background.
The Ca Ski cell line is an adherent epithelial line derived from a metastatic cervical epidermoid carcinoma in a 40-year-old female. Critically, these cells contain integrated HPV-16 and HPV-18 genomes, making them a premier model for HPV-associated oncogenesis. They are routinely used to study viral E6/E7-mediated transformation, cellular invasion mechanisms, and response to chemotherapeutic agents.
HCAR2 (hydroxycarboxylic acid receptor 2) functions as a G protein-coupled receptor for the short-chain fatty acid butyrate and the ketone body ??-hydroxybutyrate, as well as for niacin. Activation promotes coupling to G??i/o proteins, leading to inhibition of adenylyl cyclase and decreased intracellular cAMP, followed by AMPK phosphorylation. Concurrently, ??-arrestin-2 recruitment facilitates ERK1/2 activation and suppression of the NF-??B pathway, thereby downregulating pro-inflammatory cytokines like TNF-?? and IL-6, reducing COX-2-mediated prostaglandin synthesis, and triggering apoptosis in certain cancer cells. The receptor’s expression is transcriptionally upregulated by PPAR??.
Within the Ca Ski cervical carcinoma environment, HCAR2 signaling is hypothesized to exert anti-inflammatory and tumor-suppressive effects that may counter HPV-driven oncogenic pathways. Endogenous ligands such as butyrate, present in the cervicovaginal tract, could engage HCAR2 to inhibit NF-??B, potentially affecting viral oncogene expression and cellular survival. This knockout model permits dissection of HCAR2-specific effects from those mediated by other butyrate targets, enabling precise investigation of receptor-mediated modulation of HPV pathology.
Key applications include quantifying HCAR2-dependent signal transduction via western blotting of AMPK, ERK1/2, and NF-??B markers; assessing cAMP modulation; profiling cytokine expression by RT-qPCR; and evaluating apoptosis, proliferation, and migration in response to niacin or butyrate. The cells facilitate drug screening for HCAR2 agonists or antagonists and studies on metabolic-inflammatory crosstalk in cervical cancer. For further information, please contact Ascent Research.
