The ECH1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human liver adenocarcinoma line. This polyclonal pool, with disrupted ECH1 expression, facilitates the study of peroxisomal enoyl-CoA hydratase function in a neoplastic epithelial background without clonal bias.
SK-HEP-1 is a hepatic adenocarcinoma epithelial cell line widely used as a hepatocellular carcinoma model. It retains key metabolic features including active fatty acid oxidation and peroxisomal functions, making it suitable for exploring lipid metabolism in oncogenesis.
ECH1 encodes a peroxisomal enoyl-CoA hydratase essential for ??-oxidation of very long-chain and branched-chain fatty acids. ECH1 activity is transcriptionally regulated by the nuclear receptors PPARA and PPARG, which heterodimerize with RXRA and are modulated by HNF4A. The enzyme cooperates with ACOX1 and HSD17B4 in the peroxisomal ??-oxidation spiral, forming functional complexes with SCP2, and its substrate delivery is mediated by the ABCD3 transporter and PEX5 receptor. Loss of ECH1 impairs the degradation of long-chain fatty acids, reducing levels of acetyl-CoA, acyl-CoA, and ketone bodies, while perturbing bile acid biosynthesis and elevating reactive oxygen species. These metabolic shifts can blunt PPAR activation, as reduced fatty acid-derived ligands fail to sustain transcriptional activity.
Within SK-HEP-1 hepatocellular carcinoma cells, ECH1 knockout specifically disrupts peroxisomal lipid catabolism amid a reprogrammed cancer metabolic network. Hepatoma cells frequently upregulate lipid uptake and de novo synthesis; loss of ECH1 further skews lipid partitioning, potentially exacerbating triglyceride accumulation and lipotoxic stress akin to non-alcoholic fatty liver disease. This model thus permits dissection of how peroxisomal dysfunction intersects with oncogenic signaling, influences PPAR-dependent transcription, and modifies bile acid profiles, offering insights into metabolic vulnerabilities in liver cancer and metabolic syndrome.
Researchers can leverage this polyclonal ECH1 knockout model for mechanistic studies and drug screening. Standard gene disruption confirmation is performed by Western blotting for ECH1 and RT-qPCR of downstream peroxisomal transcripts such as ACOX1 and HSD17B4. Metabolic phenotyping can include fatty acid oxidation assays, Seahorse flux analysis, and lipidomic profiling to quantify very long-chain fatty acids and acylcarnitines. Complementary assays like bile acid profiling, ROS measurement, and peroxisomal immunofluorescence enable pathway-specific readouts. The model is also applicable to proliferation and drug sensitivity assays for compounds targeting hepatic lipid metabolism, and to hepatotoxicity evaluations where peroxisomal health is critical. For further details, please contact Ascent Research.
