The ALOX12 Knockout T-47D Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the T-47D human breast ductal carcinoma epithelial cell line. This mixed population harbors diverse ALOX12 gene disruptions, offering a versatile loss-of-function model that minimizes clonal selection bias and captures heterogeneity relevant to breast cancer research.
The T-47D host cell line, established from the pleural effusion of a patient with infiltrating ductal carcinoma, serves as a well-characterized model of estrogen receptor-positive (ER+)/progesterone receptor-positive (PR+) luminal A breast cancer. Its hormone-responsive phenotype and adherent epithelial morphology make it a robust platform for dissecting signaling pathways that intersect with endocrine regulation.
ALOX12 encodes arachidonate 12-lipoxygenase, a lipid-peroxidating enzyme that catalyzes the oxygenation of arachidonic acid to 12(S)-HPETE, which is rapidly reduced to 12(S)-HETE. The gene is transcriptionally regulated by SP1 and induced by pro-inflammatory cytokines including IL-1?? and TNF-??. Its enzymatic activity requires calcium ions and membrane phospholipid interactions. Following synthesis, 12-HETE functions as a lipid mediator by activating the G-protein-coupled receptor GPR31 and the nuclear receptor PPAR??, which in turn modulate NF-??B transcriptional activity and integrin-dependent adhesion. Moreover, ALOX12 promotes ferroptotic cell death by propagating lipid peroxidation, thereby integrating lipoxygenase activity with regulated necrosis and inflammatory signaling.
In T-47D cells, ALOX12-generated eicosanoids may influence estrogen-dependent proliferation, apoptosis, and metastatic potential, while its role in ferroptosis links lipid metabolism to treatment sensitivity. This polyclonal knockout model enables investigation of 12-lipoxygenase contributions to hormone-driven growth and inflammatory crosstalk within the tumor microenvironment. This flexible knockout tool allows dissection of ALOX12-dependent signaling in the context of the ER+/PR+ background, enabling exploration of cross-talk between lipid mediators and hormone receptors.
Applications include Western blot and RT-qPCR confirmation of ALOX12 ablation, LC-MS measurement of 12-HETE to verify functional knockout, and functional assays such as ferroptosis induction, lipid peroxidation quantification, cell proliferation and apoptosis analyses, and migration/invasion studies. This model supports research into arachidonic acid metabolism, eicosanoid signaling, and ferroptosis, and facilitates drug target validation in breast cancer and related diseases. For additional details or custom requests, contact Ascent Research.
