The ADAM8 Knockout U2OS Cell Line is a CRISPR/Cas9-edited knockout cell line for functional studies of ADAM8 in a human osteosarcoma epithelial system. Disruption of the ADAM8 gene ablates the full-length protein, enabling precise interrogation of its metalloproteinase and disintegrin domain activities, including ectodomain shedding, cell migration, and downstream signaling, without interference from endogenous ADAM8.
The U2OS host cell line is derived from a moderately differentiated osteosarcoma of the tibia and maintains wild-type p53 and RB tumor suppressors, offering a genetically stable platform for cancer biology research. As an adherent epithelial line widely used in the field, U2OS supports reproducible assays in migration, invasion, and proliferation, and its bone origin makes it particularly valuable for studying tumor-bone microenvironment interactions and osteosarcoma metastasis.
ADAM8 is a transmembrane metalloproteinase with a disintegrin domain that sheds ectodomains of cytokines, growth factors, and adhesion molecules. It is transcriptionally regulated by TNF-??, IL-1??, EGF, AP-1, NF-??B, and HIF-1??. Cleavage of substrates including HB-EGF, Amphiregulin, Delta-like 1, L-selectin, TNF-??, and VCAM-1 activates EGFR and Notch pathways. ADAM8 also interacts with integrins ??9??1 and ??V??3, tetraspanin CD9, fibronectin, and kinases Src and FAK. Key signaling cascades include EGFR-Ras-Raf-MEK-ERK downstream of HB-EGF shedding, and Notch intracellular domain activation of Hes/Hey transcription factors.
In U2OS cells, ADAM8-driven shedding promotes autocrine/paracrine EGFR and Notch signaling, which enhances migration and invasion??critical for osteosarcoma malignancy. The knockout line allows researchers to separate ADAM8 functions from those of related sheddases like ADAM10 and ADAM17. Given the bone-derived host, this model is ideal for examining ADAM8’s contribution to the bone microenvironment, osteoclast/osteoblast crosstalk, and metastatic niche formation, with relevance to cancers that metastasize to bone and inflammatory conditions where ADAM8 is upregulated.
Applications include osteosarcoma metastasis assays using Transwell chambers; small molecule inhibitor screening with readouts such as phospho-EGFR western blot or HB-EGF ELISA; substrate identification via proteomics; immune evasion studies by flow cytometry for L-selectin or VCAM-1; and transcriptomic analysis by RNA-seq. Validation of knockout and pathway modulation is achieved by RT-qPCR and western blotting. This cell line offers a versatile tool for ADAM8-focused research in oncology and beyond. For further information, contact Ascent Research.
