Adora2b Knockout LLC Cell Line

The Adora2b Knockout LLC Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the mouse Lewis lung carcinoma (LLC) cell line, engineered to disrupt the Adora2b gene. This product provides a stable loss-of-function model for investigating the role of the adenosine A2B receptor in lung adenocarcinoma biology. The knockout cell line is supplied as a validated cell population suitable for functional assays in cancer, immunology, and signaling research, enabling dissection of adenosine-mediated pathways in a well-characterized NSCLC model.

The parental LLC cell line is a highly aggressive lung adenocarcinoma derived from spontaneous lung tumors in C57BL/6 mice, widely used as a syngeneic model for non-small cell lung cancer (NSCLC). LLC cells exhibit rapid tumor growth, high metastatic potential, and a tumor microenvironment that recapitulates features of human NSCLC, including immune cell infiltration and hypoxic regions. They respond to adenosine signaling, making them an ideal host for studying the Adora2b receptor in a pathophysiologically relevant context.

Adora2b encodes a G protein-coupled receptor that binds extracellular adenosine and primarily couples to G??s, activating adenylate cyclase to elevate intracellular cAMP. This stimulates protein kinase A (PKA) and subsequent phosphorylation of CREB, driving transcription of proliferation and immune regulatory genes. Adora2b signaling in LLC cells is activated by adenosine from the tumor microenvironment, under the control of hypoxia/HIF-1?? and inflammatory mediators TNF-?? and IL-1??. Downstream effectors include ERK1/2, AKT, VEGF, and IL-6, connecting the receptor to MAPK/ERK, PI3K/AKT, and HIF-1?? signaling. The receptor also interacts with ??-arrestin-2 and G??q/11, though G??s?CcAMP?CPKA is the predominant cascade.

Disruption of Adora2b in LLC cells abrogates adenosine-mediated Gs signaling, leading to reduced cAMP production and impaired PKA/CREB activation, thereby impairing tumor cell proliferation, survival, and the capacity to establish an immunosuppressive microenvironment. Consequently, knockout cells may display reduced secretion of VEGF and IL-6, diminished activation of ERK1/2 and AKT, and altered responses to hypoxia. This engineered model therefore provides a powerful platform for studying the adenosine?CHIF-1?? regulatory loop, investigating mechanisms of immune evasion, and testing combination therapies with immune checkpoint inhibitors in a syngeneic NSCLC background.

The Adora2b Knockout LLC Cell Line supports a broad range of applications, including RT-qPCR and western blotting for Adora2b ablation confirmation, cAMP assays and phospho-CREB immunofluorescence for signaling analysis, and functional studies such as proliferation (MTT/CCK-8), migration/invasion (transwell), and xenograft tumor growth. Co-culture with immune cells and flow cytometry assess microenvironmental changes, while hypoxia chamber experiments investigate the adenosine?CHIF-1?? crosstalk, and ELISA for VEGF and IL-6 quantifies downstream pathway outputs. This knockout cell line is a critical tool for drug target validation and metastasis research. For additional information or customized services, please contact Ascent Research.