The EHHADH knockout NCI-H1975 polyclonal cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human NCI-H1975 lung adenocarcinoma cell line. This product features targeted disruption of the EHHADH gene, which encodes the peroxisomal bifunctional enzyme. The knockout has been generated using CRISPR/Cas9-mediated gene targeting, creating a heterogeneous population with loss-of-function mutations. This polyclonal format enables functional studies of EHHADH deficiency in a relevant cancer model for investigating peroxisomal fatty acid oxidation in lung adenocarcinoma.
The NCI-H1975 cell line is a widely used model of non-small cell lung cancer (NSCLC), derived from pleural effusion of a lung adenocarcinoma patient. This epithelial line harbors EGFR L858R and T790M mutations, conferring sensitivity to tyrosine kinase inhibitors. NCI-H1975 cells are employed in studies of EGFR signaling, drug resistance, and metabolic adaptations. EHHADH knockout in this background provides a platform to dissect the interplay between oncogenic signaling and peroxisomal lipid metabolism.
EHHADH is a peroxisomal bifunctional enzyme catalyzing hydration and dehydrogenation steps in very long-chain fatty acid ??-oxidation and participating in bile acid synthesis. It is transcriptionally regulated by PPARA in concert with PPARGC1A. EHHADH processes very long-chain acyl-CoA intermediates to generate acetyl-CoA and bile acid metabolites. It interacts with peroxisomal proteins PEX5, ACOX1, HSD17B4, and SCP2. Disruption impairs fatty acid oxidation and bile acid synthesis, disrupting lipid homeostasis.
In EGFR-mutated NCI-H1975 cells, EHHADH knockout permits investigation of peroxisomal fatty acid oxidation in NSCLC. Cancer cells often rewire metabolism; loss of EHHADH may shift reliance to glycolysis or mitochondrial oxidation. This model is relevant for studying crosstalk between PPARA/PPARGC1A and EGFR signaling in lipid metabolism, and for exploring peroxisomal disorder mechanisms in a cancer context.
These cells are suitable for Western blotting, RT-qPCR for PPARA targets, fatty acid oxidation assays, and LC-MS lipidomics to quantify very long-chain fatty acids and bile acids. Seahorse metabolic flux analysis can assess bioenergetic changes. The model enables compound screening targeting lipid metabolism in cancer. For further information, contact Ascent Research.
