The ALOX12 Knockout MCF-7 Polyclonal Cells product consists of an MCF-7 cell pool engineered by CRISPR/Cas9 to disrupt the ALOX12 gene, generating a polyclonal knockout population. This heterogeneous loss-of-function model enables studies of 12-lipoxygenase biology in a well-characterized breast adenocarcinoma background.
MCF-7 is a widely used human breast cancer cell line originally isolated from the pleural effusion of a 69-year-old Caucasian female with metastatic adenocarcinoma. These cells exhibit an adherent epithelial morphology and represent a Luminal A, estrogen receptor-positive subtype, making them a standard model for hormone-responsive breast cancer. The parental line serves as a benchmark for investigating oncogenic signaling, drug response, and metastasis.
ALOX12 encodes arachidonate 12-lipoxygenase, a lipoxygenase family member that catalyzes the oxygenation of arachidonic acid to 12(S)-hydroperoxyeicosatetraenoic acid (12(S)-HPETE), which is rapidly reduced to 12-hydroxyeicosatetraenoic acid (12-HETE). 12-HETE serves as a lipid mediator, primarily through G protein-coupled receptor GPR31, activating the MAPK (ERK1/2) and PI3K/AKT pathways. Upstream regulators include EGF, IL-6, TNF-alpha, and transcription factors SP1, NF-kB, and p53, while interacting proteins include PLA2G4A, ALOX5, and ALOX5AP. Key downstream targets are Bcl-2, Bax, MMP-2, MMP-9, VEGF, AKT, and ERK1/2. Through these interactions, ALOX12 regulates cell proliferation, survival, migration, oxidative stress, and ferroptosis.
Disruption of ALOX12 in the ER-positive MCF-7 line provides a powerful tool to dissect the contribution of 12-lipoxygenase to breast cancer progression. In this cellular context, loss of 12-HETE production can alter MAPK and PI3K/AKT signal intensity, impacting proliferation and anti-apoptotic programs. Additionally, ALOX12’s role in lipid peroxidation positions this model for investigating ferroptosis susceptibility and the interplay between eicosanoid metabolism and iron-dependent cell death. The polyclonal nature of the knockout population allows assessment of heterogeneous responses, avoiding artifacts from clonal selection while supporting robust functional genomics studies.
This polyclonal ALOX12 knockout MCF-7 pool supports diverse research applications, including studies of arachidonic acid metabolism, ferroptosis mechanisms, lipoxygenase inhibitor screening, and 12-HETE-dependent cancer cell migration. Compatible assays include Western blotting and RT-qPCR for knockout confirmation, 12-HETE quantification by ELISA or LC-MS, MTS and Annexin V assays for proliferation and apoptosis, transwell migration assays, ferroptosis induction (e.g., erastin/RSL3) with lipid peroxidation measurements, and phospho-kinase arrays for pathway profiling. RNA-seq can elucidate transcriptomic changes from ALOX12 loss. For more information, contact Ascent Research.
