ACSL4 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-mediated gene disruption model targeting the ACSL4 gene in the CAL-27 oral squamous cell carcinoma cell line. This polyclonal knockout cell population offers a heterogeneous pool of ACSL4-null cells for rigorous functional studies, minimizing clonal selection artifacts. By abolishing ACSL4 expression, this model enables dissection of its enzymatic role in PUFA activation and downstream lipid peroxide accumulation. The system is optimized for investigating ACSL4??s role in ferroptosis execution and polyunsaturated fatty acid metabolism.
The CAL-27 cell line originates from a poorly differentiated human tongue squamous cell carcinoma and expresses mutant TP53. It is a widely employed model for oral cancer biology, exhibiting aggressive proliferation, migration, and invasion. This line serves as a robust platform for evaluating molecular mechanisms and therapeutic responses in head and neck squamous cell carcinoma, particularly in studies of tumor metastasis and drug resistance.
ACSL4 catalyzes the activation of long-chain PUFAs to acyl-CoA for incorporation into membrane phospholipids, a prerequisite for ferroptosis execution through lipid peroxidation by ALOX5 and ALOX15. Its expression is regulated by SREBF1 and PPAR??, with additional modulation by TP53 and HIF1A. ACSL4 activity is functionally opposed by GPX4, which reduces lipid peroxides. The enzyme cooperates with LPCAT3 in phospholipid remodeling, thereby determining cellular sensitivity to ferroptosis inducers.
Given the relevance of ferroptosis evasion and lipid metabolic dysregulation in oral squamous cell carcinoma, ACSL4 knockout in CAL-27 cells enables dissection of ferroptotic pathways in a mutant p53 background. This model allows direct comparison of lipid peroxidation dynamics, membrane phospholipid composition, and susceptibility to ferroptosis-inducing agents such as RSL3 and erastin, providing insight into p53-dependent and -independent ferroptosis regulation.
Researchers can confirm ACSL4 knockout by Western blotting and RT-qPCR, assess lipid peroxidation via C11-BODIPY or MDA assays, and measure lipid ROS by flow cytometry. Cell viability assays with RSL3 or erastin test ferroptosis sensitivity, while LC-MS phospholipid profiling and functional assays (colony formation, migration) reveal phenotypic consequences. Its polyclonal nature maintains cellular heterogeneity, making it well-suited for drug screening campaigns and validation of lipid metabolism or ferroptosis targets. For further details, contact Ascent Research.
