The AIFM2 Knockout UM-UC-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the UM-UC-3 human urinary bladder transitional cell carcinoma epithelial cell line. These polyclonal knockout cells harbor targeted disruptions in the AIFM2 gene, also known as ferroptosis suppressor protein 1 (FSP1), providing a heterogeneous loss-of-function model for investigating ferroptosis and apoptosis pathways without single-cell cloning.
UM-UC-3 cells originate from a primary transitional cell carcinoma of the urinary bladder in a male patient and serve as a widely used in vitro model for bladder cancer research. This epithelial cell line recapitulates key features of high-grade urothelial carcinoma, including tumorigenesis, invasive potential, and drug response, offering a clinically relevant background for studying AIFM2-dependent mechanisms in bladder cancer.
AIFM2 encodes a flavoprotein oxidoreductase that catalyzes the reduction of coenzyme Q10 (CoQ10) to ubiquinol (CoQ10H2), a lipophilic antioxidant that suppresses ferroptosis by inhibiting lipid peroxidation. It also participates in caspase-independent apoptosis. Activity is regulated by upstream signals including p53 and NRF2 under oxidative stress and requires cofactors FAD and NAD(P)H. Downstream, AIFM2-generated CoQ10H2 attenuates lipid peroxidation, preventing ferroptotic death. Within the ferroptosis network, AIFM2 functions alongside GPX4 and SLC7A11, opposing the pro-ferroptotic action of ACSL4, integrating CoQ10 metabolism into redox homeostasis.
CRISPR-mediated ablation of AIFM2 in UM-UC-3 cells eliminates FSP1-dependent ferroptosis protection, drastically sensitizing cells to ferroptosis inducers such as erastin and RSL3 and leading to increased lipid peroxidation. Additionally, disruption of AIFM2 may alter caspase-independent apoptotic responses under oxidative stress, influencing cell fate. Given the role of ferroptosis evasion in drug resistance, this knockout model is invaluable for deciphering how bladder cancer cells escape oxidative death and for testing strategies to restore ferroptosis sensitivity.
Research applications encompass ferroptosis mechanism studies using lipid peroxidation probes like C11-BODIPY, cell viability assays, and expression analysis via western blotting and RT-qPCR. The model supports screening of ferroptosis-inducing anticancer compounds, coenzyme Q10 metabolite profiling, caspase-independent apoptosis assays, and colony formation assays. It enables interrogation of p53/NRF2-regulated antioxidant responses and the interplay between AIFM2, GPX4, SLC7A11, and ACSL4 in bladder cancer pathogenesis. For further details, please contact Ascent Research.
