The ALOX12 Knockout SK-OV-3 Polyclonal Cells constitute a CRISPR/Cas9-mediated polyclonal knockout population in which the human ALOX12 gene (encoding arachidonate 12-lipoxygenase) has been disrupted in the SK-OV-3 ovarian adenocarcinoma cell line. Supplied as an unselected pool, these polyclonal knockout cells offer a genetically heterogeneous loss-of-function model for studying ALOX12-dependent processes without clonal artifacts. The editing targets the endogenous ALOX12 locus, creating a cellular reagent suitable for pathway analysis and compound evaluation in a disease-relevant host.
The host SK-OV-3 cell line is a well-characterized model of human ovarian adenocarcinoma, originally established from the ascites of a patient. These cells display epithelial morphology, express HER2/neu and mutant TP53, and are tumorigenic in immunodeficient mice. SK-OV-3 is widely employed to study ovarian cancer tumorigenesis, metastasis, and chemoresistance, particularly to platinum-based agents and taxanes. The line??s intrinsic molecular features make it a relevant platform for interrogating ALOX12 function within a tumorigenic context.
ALOX12 catalyzes the conversion of arachidonic acid, liberated by cPLA2, into 12(S)-hydroperoxyeicosatetraenoic acid (12-HPETE), which is rapidly reduced to 12(S)-HETE. The bioactive lipid 12-HETE signals through the G-protein?coupled receptor GPR31 to activate downstream PI3K/AKT and ERK pathways, thereby promoting cell proliferation, survival, and migration. Upstream, ALOX12 expression is transcriptionally regulated by SP1, AP-1, and NF-??B in response to growth factors including EGF and TGF???. The enzyme collaborates with integrin ??4 and requires membrane association and calcium for full activity, linking integrin signaling to eicosanoid production.
In the SK-OV-3 ovarian cancer context, ALOX12-derived 12-HETE drives PI3K/AKT and ERK signaling, contributing to an aggressive phenotype characterized by enhanced proliferation, migration, and resistance to cisplatin and paclitaxel. The mutant TP53 background may further potentiate ALOX12-mediated tumorigenic signaling. Disruption of ALOX12 in this cell line provides a valuable loss-of-function model for dissecting 12-lipoxygenase-dependent mechanisms of ovarian cancer progression, including epithelial-mesenchymal transition, integrin activation, and focal adhesion dynamics. The polyclonal nature of the knockout population minimizes clonal bias, making it suitable for reproducible functional genomics and drug sensitivity profiling.
These polyclonal knockout cells enable detailed investigation of ALOX12??s role in ovarian cancer proliferation, migration, and chemoresistance. Researchers can perform MTT or IncuCyte proliferation assays, Transwell migration/invasion assays, and drug sensitivity testing with cisplatin or paclitaxel. Signaling pathway analysis via Western blotting for phospho-AKT and phospho-ERK, along with RT?qPCR, complements lipidomic quantification of 12-HETE. The model is also suited for 12-LOX inhibitor screening and in vivo xenograft studies. For further information, please contact Ascent Research.
