The ECHDC3 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal population derived from the human hepatic adenocarcinoma cell line SK-HEP-1. This loss-of-function model enables the investigation of ECHDC3 (enoyl-CoA hydratase domain-containing 3) in a hepatocellular carcinoma background. The polyclonal nature of this knockout product provides a heterogeneous pool of edited cells, reflecting a range of gene-disruption events and facilitating population-level studies of metabolic and signaling disruptions without clonal selection bias.
The parental SK-HEP-1 cell line, originally established from the ascites of a patient with liver adenocarcinoma, is widely used as an in vitro model for hepatocellular carcinoma (HCC). SK-HEP-1 cells retain many metabolic features of hepatic tissue, including active fatty acid oxidation and amino acid catabolism, making them particularly suitable for studying lipid metabolism and its dysregulation in liver cancer. Their mesenchymal-like phenotype also supports assays related to cancer cell migration and invasion in the context of metabolic stress.
ECHDC3 encodes a mitochondrial enoyl-CoA hydratase that catalyzes the hydration of trans-2-enoyl-CoA to 3-hydroxyacyl-CoA in the beta-oxidation pathway. This reaction is essential for the stepwise degradation of long-chain fatty acids, linking fatty acid degradation to the valine, leucine, and isoleucine degradation pathways. ECHDC3 activity is transcriptionally regulated by PPAR-alpha, PGC-1alpha, and HNF4-alpha, and its enzymatic function directly produces 3-hydroxyacyl-CoA and contributes to acetyl-CoA, NADH, and FADH2 generation. The enzyme interacts with the mitochondrial trifunctional protein subunits HADHA and HADHB, as well as electron transfer flavoproteins ETF and ETFDH, underscoring its integration into the broader beta-oxidation machinery.
In SK-HEP-1 hepatocellular carcinoma cells, ECHDC3 disruption is anticipated to impair mitochondrial fatty acid breakdown, leading to the accumulation of enoyl-CoA intermediates and a reduction in acetyl-CoA production. This metabolic bottleneck may compromise ATP synthesis and redox balance, thereby sensitizing cells to metabolic stress and potentially unmasking vulnerabilities relevant to HCC biology. Given the central role of lipid metabolism in liver physiology and pathology, this knockout model offers a powerful tool to dissect how defective beta-oxidation influences tumor cell survival, proliferation, and response to nutrient-limited microenvironments.
Researchers can employ this ECHDC3 polyclonal knockout pool in a wide array of experimental contexts, including metabolic flux analyses with [13C]palmitate tracing to quantify beta-oxidation impairments, Seahorse XF assays to assess mitochondrial respiration, and lipid droplet visualization via Oil Red O staining. Downstream applications encompass transcriptomic profiling by RNA-seq, protein-level validation by Western blotting, and functional studies such as ATP measurement and apoptosis assays under lipid-rich or nutrient-deprived conditions. These cells are particularly suited for drug sensitivity screens targeting metabolic vulnerabilities in HCC. For further inquiries or to discuss custom projects, please contact Ascent Research.
