The ALOX12 Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal human AGS cell population harboring targeted disruption of the ALOX12 gene. This model ablates expression of arachidonate 12-lipoxygenase, the enzyme responsible for converting arachidonic acid into the eicosanoid mediator 12-HETE. By eliminating 12-HETE production, the system enables detailed investigation of 12-lipoxygenase-driven signaling in gastric cancer without relying on pharmacological inhibition.
The host AGS line is a widely used human gastric adenocarcinoma epithelial cell model derived from a 54-year-old Caucasian female. AGS cells recapitulate key hallmarks of gastric carcinogenesis and are routinely applied in studies of tumor cell proliferation, apoptosis, and invasion. Their defined genetic background and stable in vitro growth make them an ideal platform for gene-editing approaches aimed at dissecting molecular mechanisms of gastric cancer.
ALOX12 functions within the arachidonic acid metabolic pathway, generating 12-HETE that signals through the G protein-coupled receptor GPR31. Downstream, GPR31 engagement stimulates the PI3K-Akt and MAPK/ERK1/2 cascades, promoting Bcl-2-mediated survival and cell cycle progression while suppressing pro-apoptotic Bax and Caspase-3. ALOX12 expression is induced by cytokines IL-4, IL-13, and TNF-?? via STAT6 and NF-??B, and its catalytic activity depends on calcium and phospholipase A2-mediated release of arachidonic acid from membrane phospholipids. Thus, ALOX12 knockout severs a conduit connecting inflammatory signals to proliferative and anti-apoptotic effectors.
In AGS cells, the ALOX12/12-HETE/GPR31 axis sustains oncogenic signaling, and its disruption leads to diminished ERK1/2 and Akt phosphorylation, reduced colony formation, and enhanced apoptosis. This polyclonal knockout model therefore offers a relevant tool to investigate how eicosanoid dysregulation contributes to gastric adenocarcinoma pathology and to test strategies that interfere with 12-HETE production or GPR31 function.
Typical applications include MTT and colony formation assays to assess proliferation, Annexin V/PI staining and Caspase-3 activation assays for apoptosis, and Transwell/Matrigel assays for migration and invasion. 12-HETE ELISA verifies loss of metabolite, while western blot for ALOX12, phospho-ERK1/2, and phospho-Akt confirms pathway attenuation. The cells are also amenable to transcriptomic analysis by RNA-seq and to drug target validation studies. For additional details, please contact Ascent Research.
