The ALOX12 Knockout Ca Ski Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Ca Ski human cervical carcinoma cell line, engineered to disrupt ALOX12 gene expression. This polyclonal cell product offers a loss-of-function model for studying arachidonate 12-lipoxygenase function in oncogenic and inflammatory signaling. The CRISPR/Cas9-mediated gene disruption avoids clonal selection, yielding a heterogeneous pool that retains the parental Ca Ski genomic background. It is intended for biomedical research into cancer biology, inflammation, and lipid signaling.
The Ca Ski line is an epithelial cell model from a cervical epidermoid carcinoma, carrying integrated HPV-16 genome and constitutively expressing E6 and E7 oncoproteins. Widely used for HPV-related carcinogenesis studies, these cells exhibit epithelial morphology and active tumor-relevant signaling pathways. Combining ALOX12 knockout with this HPV-positive background enables dissection of crosstalk between viral oncogenesis and eicosanoid signaling in cervical cancer.
ALOX12 encodes arachidonate 12-lipoxygenase, which catalyzes conversion of arachidonic acid to 12(S)-HPETE, then reduced to 12(S)-HETE. 12-HETE binds the GPR31 receptor, activating PI3K/AKT signaling to promote cell proliferation and migration via integrin activation. ALOX12 expression is regulated by cytokines such as IL-4 and growth factors, engaging transcription factors SP1 and AP-2 through MAPK cascades. It also interacts with membrane phospholipids and peroxisome proliferator-activated receptors. Knockout of ALOX12 in Ca Ski cells disrupts 12-HETE production, uncoupling GPR31-mediated PI3K/AKT phosphorylation and attenuating downstream oncogenic signals.
In cervical carcinoma, ALOX12 may drive Ca Ski cell aggressiveness through autocrine 12-HETE signaling that sustains proliferation and invasion. HPV-16 E6/E7 oncoproteins subvert host pathways, and loss of ALOX12 allows testing of cooperative roles between viral transformation and lipid mediator synthesis. The knockout model enables interrogation of PI3K/AKT dependency downstream of GPR31 and the consequences of eicosanoid synthesis interruption. The polyclonal population mimics tumor heterogeneity, supporting studies on adaptive resistance and response diversity following pathway disruption.
These polyclonal knockout cells are suitable for investigating 12-HETE functions using Western blotting, RT-qPCR, 12-HETE ELISA, and phospho-AKT analysis. Functional assays may include migration, invasion, and cell viability measurements, complemented by lipidomics profiling. Research applications encompass elucidating ALOX12 contribution to cervical cancer progression, evaluating 12-lipoxygenase as a therapeutic target, and exploring lipid signaling in HPV-driven neoplasia. For further information, please contact Ascent Research.
