The ACSL4 Knockout HGC-27 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population derived from the HGC-27 human gastric carcinoma cell line, in which the ACSL4 gene has been disrupted via CRISPR/Cas9-mediated gene editing. This polyclonal knockout cell pool provides a loss-of-function model for investigating ACSL4-dependent processes, particularly in the context of ferroptosis and lipid metabolism. The heterogeneous nature of the polyclonal population reflects a range of editing outcomes, making it suitable for studying gene function without the limitations of clonal selection.
HGC-27 cells originate from a poorly differentiated gastric adenocarcinoma and were established from a lymph node metastasis, retaining epithelial morphology. This cell line is widely used in gastric cancer research to model metastatic behavior, treatment resistance, and metabolic reprogramming. As a gastric carcinoma line derived from a metastatic site, HGC-27 cells exhibit aggressive growth characteristics and are responsive to ferroptosis induction, making them a relevant model system for ACSL4 functional studies.
ACSL4 (acyl-CoA synthetase long-chain family member 4) catalyzes the activation of long-chain polyunsaturated fatty acids, particularly arachidonic acid, to form arachidonoyl-CoA. This reaction is a critical step in the incorporation of these fatty acids into membrane phospholipids, a process facilitated by lysophosphatidylcholine acyltransferase 3 (LPCAT3). Enriched PUFA-phospholipids serve as substrates for lipoxygenases such as ALOX15, leading to lipid peroxidation and the execution of ferroptotic cell death, counterbalanced by the glutathione-dependent peroxidase GPX4. ACSL4 is transcriptionally regulated by NRF2, PPAR??, and SREBP1, while its activity can be influenced by endoplasmic reticulum stress and p53 signaling. The ACSL4/LPCAT3/ALOX15/GPX4 axis constitutes the core machinery of ferroptosis execution, with ACSL4 acting as a key determinant of cellular sensitivity to this non-apoptotic cell death pathway.
In HGC-27 cells, ACSL4 expression contributes to the intrinsic susceptibility to ferroptosis. Knockout of ACSL4 in this cell line disrupts the generation of oxidizable phospholipid species, thereby conferring resistance to ferroptosis inducers such as erastin and RSL3. This model also perturbs arachidonic acid metabolism and phospholipid remodeling, potentially affecting downstream pathways like pro-inflammatory eicosanoid production and PPAR signaling. By ablating ACSL4, researchers can dissect the interconnection between lipid metabolism reprogramming and cell death mechanisms in a gastric carcinoma background, which is highly relevant for understanding ferroptosis resistance in aggressive gastric cancer.
This ACSL4 knockout cell product is employed in a variety of experimental workflows, including lipid peroxidation measurement using C11-BODIPY, ferroptosis sensitivity profiling, and metabolomic analyses to track altered fatty acid utilization. Researchers can combine this model with drug screening campaigns to identify novel ferroptosis inducers, or utilize cell migration and invasion assays to investigate the role of ACSL4 in gastric cancer metastasis. Additional applications include xenograft tumor studies to assess the impact of ACSL4 loss on tumor growth and treatment response in vivo. CRISPR on-target editing validation by western blotting and RT-qPCR is recommended to confirm gene disruption. For additional information or personalized technical support, please contact Ascent Research.
