The ALOX12 Knockout 143B Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human ALOX12 gene in the 143B osteosarcoma cell line. This product provides a heterogeneous pool of cells carrying diverse loss-of-function mutations, avoiding clonal selection artifacts while maintaining genetic diversity for pooled functional assays, drug screening, and metastasis research. The knockout model serves as a versatile tool to investigate arachidonic acid metabolism, lipid signaling, and ferroptosis regulation in a clinically relevant bone cancer context.
The parental 143B line is a well-characterized human osteosarcoma model with a highly metastatic phenotype and mesenchymal origin. Derived from the HOS (TE-85) cell line, 143B cells rapidly proliferate, exhibit invasive behavior, and form lung metastases in xenograft models, recapitulating aggressive disease features. This background enables genetic dissection of molecular drivers of osteosarcoma progression, making it ideal for assessing the contribution of 12-lipoxygenase to malignant properties.
ALOX12 encodes arachidonate 12-lipoxygenase, which catalyzes the dioxygenation of arachidonic acid to 12-hydroperoxyeicosatetraenoic acid (12-HPETE), rapidly reduced to the potent lipid mediator 12-hydroxyeicosatetraenoic acid (12-HETE). This reaction occurs downstream of phospholipase A2-mediated membrane phospholipid hydrolysis. The gene is transcriptionally regulated by p53, TNF-alpha, IL-1beta, NF-kB, and AP-1. 12-HETE signals through GPR31 and PPARgamma, activating MAPK cascades (ERK, p38, JNK) and modulating caspase-dependent apoptosis and lipid peroxidation. The enzyme’s pro-ferroptotic activity is counterbalanced by GPX4, placing ALOX12 at a key node linking proliferation, apoptosis, and ferroptosis in platelet biology and cancer.
In the 143B osteosarcoma background, ALOX12-derived 12-HETE has been implicated in promoting tumor cell proliferation, survival, migration, and metastasis, while also participating in p53-mediated apoptotic responses under genotoxic stress. The knockout model therefore permits separation of 12-lipoxygenase-dependent versus independent tumorigenic processes, with particular relevance to ferroptosis sensitivity and metastatic signaling. The polyclonal format reflects tumor heterogeneity and facilitates identification of robust ALOX12-dependent phenotypes in cancer.
Researchers can employ this product for applications including Western blotting and RT-qPCR to verify knockout efficiency, LC-MS to confirm loss of 12-HETE production, and cell proliferation (MTT/CCK-8) or Transwell migration/invasion assays to assess functional impact. Ferroptosis induction with Erastin or RSL3, combined with lipid ROS detection, allows investigation of ferroptosis mechanisms, while apoptosis analysis (Annexin V/PI) and xenograft tumor growth studies evaluate in vivo tumorigenicity. These approaches support research in osteosarcoma biology, colorectal and prostate cancer, platelet dysfunction, and atherosclerosis. For further information, please contact Ascent Research.
