The HCAR2 Knockout 143B Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal cell population derived from the human 143B osteosarcoma cell line, engineered for targeted disruption of the HCAR2 gene. This loss-of-function model enables investigation of HCAR2-mediated signaling pathways in a bone cancer context without altering the intrinsic genetic background of the host cells. The polyclonal format provides a heterogeneous knockout pool, avoiding clonal selection bias and preserving population-level biological variability, which is advantageous for studying gene function in a more physiologically relevant cellular context.
The 143B cell line is a subclone of the HOS osteosarcoma line, characterized by KRAS and TP53 mutations, and exhibits an epithelial-like morphology with highly aggressive and metastatic properties. Widely employed as a model for human osteosarcoma, 143B cells recapitulate key features of malignant bone tumors, including rapid proliferation, invasiveness, and the capacity to form metastases in vivo. This genetic and phenotypic background provides a clinically relevant platform for examining the role of metabolic and inflammatory regulators such as HCAR2 in tumor progression.
HCAR2 encodes a Gi-coupled receptor activated by endogenous ligands including niacin, butyrate, ??-hydroxybutyrate, and acetoacetate. Upon ligand binding, HCAR2 triggers G??i-mediated inhibition of adenylate cyclase, leading to decreased intracellular cAMP levels and subsequent suppression of protein kinase A (PKA) activity. This cascade reduces hormone-sensitive lipase-mediated lipolysis and attenuates NF-??B-driven pro-inflammatory responses, while concurrently promoting AMPK-mediated anti-inflammatory effects. HCAR2 also engages ??-arrestin-2 and GRK2, and mitogenic signaling through MAPK/ERK pathways, integrating metabolic and inflammatory signals.
In the context of osteosarcoma, HCAR2 may modulate tumor cell metabolism, inflammatory microenvironment, and immune evasion. The 143B knockout model allows dissection of HCAR2-dependent effects on lipolysis, cAMP dynamics, and NF-??B activity??pathways implicated in cancer cell survival, migration, and resistance to therapy. By eliminating HCAR2 expression, researchers can assess its contribution to the aggressive behavior of osteosarcoma cells and evaluate the dependency of downstream kinases such as AMPK and MAPK/ERK on receptor activity.
Typical applications include mechanistic studies of niacin and butyrate signaling in bone cancer, metabolic regulation of tumor aggressiveness, and the role of HCAR2 in inflammation-driven carcinogenesis. This model is also suitable for screening HCAR2 agonists or antagonists, and for functional genomics approaches such as RNA-seq transcriptomics. Assays including Western blotting, RT-qPCR, cAMP accumulation, lipolysis quantification, NF-??B luciferase reporters, phospho-AMPK detection, and migration/invasion assays are readily applicable. For further information or to request a quotation, please contact Ascent Research.
