The ALOX12 Knockout UM-UC-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from human bladder transitional cell carcinoma cells, designed for loss-of-function studies of arachidonate 12-lipoxygenase (ALOX12). This polyclonal product comprises a heterogeneous pool of UM-UC-3 cells carrying diverse CRISPR/Cas9-mediated disruptions of the ALOX12 gene, providing a population-level model that avoids clonal selection artifacts. The gene disruption impairs the enzymatic conversion of arachidonic acid to 12-hydroperoxyeicosatetraenoic acid (12-HPETE) and its reduced product 12-hydroxyeicosatetraenoic acid (12-HETE), eliminating the primary lipid mediator produced by this lipoxygenase.
UM-UC-3 is a well-characterized human urinary bladder transitional cell carcinoma cell line derived from a primary bladder carcinoma. These malignant urothelial cells are extensively used to study bladder cancer biology, including tumor cell migration, invasion, and drug response. UM-UC-3 cells exhibit activation of signaling pathways such as MAPK, NF-??B, and STAT3, which intersect with ALOX12-mediated lipid signaling, making them a physiologically relevant host for ALOX12 knockout studies.
ALOX12 catalyzes the oxygenation of arachidonic acid at carbon-12, forming 12-HPETE, which is rapidly reduced to 12-HETE. 12-HETE activates the G-protein-coupled receptor GPR31, triggering MAPK/ERK and NF-??B signaling to promote cell migration, inflammation, and tumor progression. ALOX12 expression is regulated by cytokines (IL-1??, TNF-??), growth factors (EGF, TGF-??), and hypoxia, acting through transcription factors STAT3 and NF-??B. Downstream effectors include integrin activation, small GTPases Rac and RhoA, MMP induction, and ROS production. ALOX12 functionally interacts with calcium, membrane phospholipids, integrin ??4 (ITGB4), and cPLA2.
In bladder carcinoma, ALOX12-driven 12-HETE/GPR31 signaling contributes to aggressive phenotypic traits. This polyclonal ALOX12 knockout model enables direct assessment of the 12-lipoxygenase axis in UM-UC-3 cells, facilitating dissection of its role in transmigration, invasion, and inflammation-associated malignancy. The heterogeneous knockout population better mimics tumor genetic variability than monoclonal lines, providing a robust platform for investigating the interplay between chronic inflammatory stimuli and ALOX12 signaling in urothelial cancer progression.
These cells are suited for a broad range of functional assays, including Western blotting and RT-qPCR for gene disruption validation, enzyme activity assays and 12-HETE ELISA for quantifying lipoxygenase product loss, and migration (wound healing) and invasion (transwell) assays for phenotyping. They also enable proliferation and drug sensitivity testing, RNA-seq transcriptomic profiling, and phospho-protein analysis of MAPK and NF-??B networks. Altogether, the model supports target validation, pathway dissection, and preclinical evaluation of lipoxygenase inhibitors. For further technical details or custom assay support, please contact Ascent Research.
