The HADH Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the HADH gene in the human SK-HEP-1 liver adenocarcinoma-derived cell line. This model provides a loss-of-function system for studying short-chain fatty acid metabolism and its regulatory roles. The polyclonal knockout format reflects heterogeneous gene disruption effects, suitable for functional genomics and screening applications.
SK-HEP-1 cells, isolated from liver adenocarcinoma, co-express endothelial (CD31, von Willebrand factor) and epithelial markers, serving as a hepatic endothelial model with tumorigenic properties. This hybrid phenotype enables investigations of endothelial function, angiogenesis, and tumor biology in a liver context. Combined with HADH knockout, it permits metabolic pathway analysis in a transformed endothelial-like background.
HADH (short-chain 3-hydroxyacyl-CoA dehydrogenase) catalyzes the NAD+-dependent oxidation of 3-hydroxyacyl-CoA to 3-ketoacyl-CoA, the third step of mitochondrial short-chain fatty acid ??-oxidation. This supplies acetyl-CoA for the TCA cycle and NADH for oxidative phosphorylation, linking lipid catabolism to ATP synthesis. HADH is transcriptionally regulated by PPAR?? and HNF4??, and its activity is modulated by insulin and glucagon. The enzyme homodimerizes and partners with electron transfer flavoprotein (ETF) to feed electrons into the respiratory chain. In pancreatic ??-cells, HADH controls insulin secretion by influencing the ATP/ADP ratio and redox status.
Ablation of HADH in SK-HEP-1 cells impairs short-chain fatty acid oxidation, potentially causing substrate accumulation and metabolic reprogramming. This hepatic endothelioid model is particularly valuable for dissecting lipid metabolism, ketogenesis, and mitochondrial dysfunction. Endothelial marker expression additionally allows exploration of metabolic?Cvascular crosstalk in the tumor microenvironment. The knockout can be used to assess HADH??s role in energy homeostasis, oxidative stress, and cell viability under metabolically challenged conditions.
Key applications include fatty acid oxidation assays, ATP quantification, and mitochondrial respiration analysis via Seahorse. Researchers can study HADH??s impact on insulin secretion using heterologous systems or model hyperinsulinemic hypoglycemia and HADH deficiency. Transcriptomic (RNA-seq, RT-qPCR) and proteomic (Western blot) profiling can delineate downstream pathways. The model also supports drug discovery efforts targeting metabolic vulnerabilities in liver cancer. For further details or technical consultation, contact Ascent Research.
