The ALOX12 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in the human SK-HEP-1 hepatic adenocarcinoma cell line, achieving targeted disruption of the ALOX12 gene. This knockout model provides a genetically defined system for loss-of-function studies of arachidonate 12-lipoxygenase, a key enzyme in lipid mediator biosynthesis. The polyclonal nature allows immediate use in pooled format, facilitating analysis of population-level effects without clonal selection artifacts. The product is designed for robust interrogation of ALOX12-dependent phenotypes in hepatocellular carcinoma models.
SK-HEP-1 is an immortalized human cell line originally derived from a patient with liver adenocarcinoma, exhibiting epithelial morphology and widely employed as a model for hepatocellular carcinoma (HCC). This cell line retains many characteristics of malignant hepatocytes and is commonly utilized to study tumorigenesis, metastasis, and drug responses. Its genetic and signaling background provides a clinically relevant platform for investigating the roles of arachidonic acid-metabolizing enzymes in HCC progression. The introduction of an ALOX12 knockout into this context enables dissection of 12-lipoxygenase contributions to cancer cell behavior.
ALOX12 encodes arachidonate 12-lipoxygenase, which converts arachidonic acid to 12S-HPETE and then to 12-HETE. This bioactive lipid signals through GPR31, activating downstream PI3K/AKT/mTOR and ERK1/2 cascades. The enzyme is transcriptionally regulated by upstream cytokines such as TNF??, IL-4, IL-13, and TGF?? via STAT6, NF-??B, and HIF-1??. 12-HETE promotes integrin ??1 activation, focal adhesion kinase signaling, and MMP-9 expression, and modulates Bcl-2 family proteins to influence apoptosis. Crosstalk with ALOX5 and cPLA2 integrates this axis into broader eicosanoid networks.
In SK-HEP-1 cells, ALOX12 knockout disrupts 12-HETE-driven signaling that contributes to HCC aggressiveness, including proliferation, migration, and invasion. This model allows direct assessment of tumorigenic dependency on ALOX12, particularly through PI3K/AKT and ERK hyperactivation and Bcl-2-mediated apoptosis regulation. Eliminating endogenous 12-HETE production facilitates dissection of its roles in tumor microenvironment interactions and metastatic potential.
This tool supports diverse assays including western blot for ALOX12 protein, RT-qPCR for transcript levels, LC-MS/MS for 12-HETE quantification, proliferation and Transwell migration/invasion assays, Annexin V apoptosis detection, and phospho-AKT/ERK analysis. It is suitable for drug target validation by comparing inhibitor effects to knockout phenotypes. For further details and ordering, please contact Ascent Research.
