The ECHDC1 Knockout SK-HEP-1 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human hepatic adenocarcinoma cell line SK-HEP-1. This product introduces targeted disruption of the ECHDC1 gene, which encodes a mitochondrial enoyl-CoA hydratase, to generate a loss-of-function model for studying fatty acid beta-oxidation and metabolic signaling. The polyclonal format provides a heterogeneous mixture of cells with diverse editing events, reflecting a population-level knockout effect without clonal selection. This approach is ideal for experiments where genetic heterogeneity mimics physiological conditions or when downstream assays do not require isogenic clonal lines.
The host cell line, SK-HEP-1, originates from the ascites of a 52-year-old male with liver adenocarcinoma and is widely used as a model for hepatic function and hepatocellular carcinoma research. SK-HEP-1 cells exhibit epithelial morphology and retain key metabolic features of liver-derived cells, making them a suitable platform for investigating energy metabolism and lipid processing pathways. Their adenocarcinoma background further enables studies that intersect metabolic reprogramming with oncogenic processes.
ECHDC1 functions within the mitochondrial short-chain fatty acid beta-oxidation pathway, catalyzing the hydration of enoyl-CoA intermediates to 3-hydroxyacyl-CoA. This enzyme acts in concert with other key beta-oxidation enzymes, including the interacting factors ECHS1, HADHA, HADHB, and ACADM. Upstream, ECHDC1 expression is regulated by metabolic sensors and transcription factors such as PPAR??, PGC-1??, and AMPK, which coordinate energy homeostasis. Disruption of ECHDC1 mechanistically leads to accumulation of enoyl-CoA substrates and reduced production of downstream targets acetyl-CoA, 3-hydroxyacyl-CoA, and ATP, thereby impairing cellular energy homeostasis and promoting lipid accumulation. This knockout model thus provides a tool to dissect the PPAR signaling pathway and mitochondrial energy metabolism.
Within the hepatic adenocarcinoma context of SK-HEP-1 cells, loss of ECHDC1 is particularly relevant for modeling non-alcoholic fatty liver disease (NAFLD) and hepatic steatosis, as impaired fatty acid oxidation is a hallmark of these conditions. The polyclonal knockout population allows researchers to study how heterogeneous ECHDC1 ablation affects lipid droplet formation, oxidative phosphorylation, and cancer cell viability. This model may reveal compensatory metabolic adaptations, such as upregulated glycolysis, that are common in metabolic syndrome and liver cancer.
Researchers can use these cells in functional assays, including fatty acid oxidation assays with 14C-palmitate, Seahorse flux analysis for OCR and ECAR, Oil Red O staining, ATP luminescence assays, Western blotting, and RT-qPCR of PPAR?? targets. Together, these applications enable detailed investigation of metabolic reprogramming in hepatocellular carcinoma and hepatic lipid metabolism. For additional technical information and ordering, please contact Ascent Research.
