HCAR1 Knockout MG-63 Cell Line

The HCAR1 Knockout MG-63 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from Homo sapiens MG-63 osteosarcoma cells. This product provides a defined loss-of-function model for studying HCAR1-mediated signaling by disrupting the target gene through CRISPR/Cas9-mediated gene disruption. It is supplied as a stable cell line suitable for a range of functional and molecular assays in cancer biology, signal transduction, and metabolic research.

The MG-63 host cell line originates from a human osteosarcoma and displays an osteoblast-like phenotype with epithelial morphology and capacity for bone matrix mineralization. These characteristics make MG-63 a well-established model for investigating bone tumor biology, osteoblast differentiation, and skeletal metastasis. The cell line retains responsiveness to hormonal and metabolic cues, providing a physiologically relevant background for gene knockout studies.

HCAR1 encodes a G protein-coupled receptor activated by extracellular lactate, functioning primarily through G??i/o proteins. Upon ligand binding, HCAR1 inhibits adenylyl cyclase, leading to reduced intracellular cAMP levels and diminished protein kinase A (PKA) activity. This cascade attenuates lipolysis by suppressing hormone-sensitive lipase (HSL) and adipose triglyceride lipase (ATGL) and dampens pro-inflammatory signaling through NF-??B inhibition. The receptor also engages G?¦? dimers to stimulate PI3K/Akt pathways and recruits ??-arrestin-2 and GRK2, which can activate MAPK/ERK signaling. Upstream regulators include hypoxia, insulin, exercise, and inflammatory cytokines such as TNF-?? and IL-1??, situating HCAR1 at the intersection of metabolic and immune responses.

In the MG-63 osteosarcoma context, HCAR1 likely integrates microenvironmental lactate signals with tumor cell behavior. Elevated lactate, a hallmark of glycolytic tumors, may promote osteosarcoma progression, migration, and bone metastasis through HCAR1-dependent pathways. This knockout cell line enables dissection of HCAR1??s contribution to tumor-stroma interactions, lactate sensing, and osteosarcoma malignancy without confounding endogenous receptor activity.

Typical research applications include cancer metabolism studies, tumor microenvironment investigations, lactate signaling in bone metastasis, immunometabolism, and drug target validation. The knockout cell line is compatible with assays such as cAMP accumulation measurement, Western blotting for phospho-PKA substrates, glycerol release lipolysis assays, Transwell migration/invasion, RT-qPCR for target gene expression, co-immunoprecipitation of G??i interactions, and lactate dose-response experiments. For further technical details and ordering, please contact Ascent Research.