This product provides a CRISPR/Cas9-edited polyclonal knockout cell population of SK-HEP-1 cells carrying targeted disruption of the HADHB gene. The polyclonal format offers a heterogeneous pool of knockout cells without single-cell cloning, ensuring that the loss-of-function phenotype is robustly represented across the population. This model is designed for investigating HADHB-dependent biological processes and metabolic pathways in a pooled context, making it suitable for studies where clonal variation is not desired.
The SK-HEP-1 host cell line is a human liver adenocarcinoma line with endothelial-like properties, widely utilized as a hepatocellular carcinoma (HCC) model. These cells display a unique blend of hepatic and endothelial features, which makes them particularly useful for studying tumor cell metabolism, angiogenesis, and the tumor microenvironment. Their derivation from a liver adenocarcinoma situates them as a relevant cellular context for dissecting metabolic reprogramming in HCC.
HADHB encodes the beta subunit of the mitochondrial trifunctional protein (MTP), which catalyzes the hydration, dehydrogenation, and thiolysis steps of long-chain fatty acid beta-oxidation. This enzymatic activity produces acetyl-CoA, NADH, and FADH2, which feed into the tricarboxylic acid cycle and oxidative phosphorylation. The expression of HADHB is transcriptionally regulated by PPARA and PPARG, and its activity is modulated by AMPK and PGC1A in response to cellular energy status. HADHB functions as part of the MTP complex, physically associating with HADHA, and interacts with other beta-oxidation enzymes such as ECHS1 and ACAA2. These interactions are integrated within the broader fatty acid catabolism network, which includes CPT1A, CPT2, ACADVL, and ACADM. Disruption of HADHB therefore leads to a block in long-chain fatty acid utilization, accumulation of long-chain acylcarnitine intermediates, and reduced ketone body production, while triggering compensatory signaling through the AMPK-mTORC1 axis.
In the context of SK-HEP-1 cells, the HADHB knockout creates a compelling model of mitochondrial fatty acid oxidation deficiency. Hepatocellular carcinoma cells are known to rewire lipid metabolism to support proliferation and survival, and loss of HADHB forces metabolic adaptation, often increasing reliance on glycolysis or glutaminolysis. The resultant energetic stress and acylcarnitine buildup recapitulate metabolic signatures of mitochondrial trifunctional protein deficiency, a disorder linked to hypoglycemia, cardiomyopathy, and neurological symptoms. This model thus enables researchers to examine how defects in long-chain fatty acid oxidation reshape tumor cell bioenergetics, lipid homeostasis, and susceptibility to metabolic stress.
This polyclonal HADHB knockout cell population is suitable for a broad range of metabolic assays. Fatty acid oxidation can be assessed via Seahorse respirometry or radiolabeled tracing, acylcarnitines by LC-MS, and ATP by luminescence. Lipid accumulation is monitored with Oil Red O, while disruption is confirmed by western blotting and RT-qPCR. Viability assays under metabolic or drug challenge probe chemoresistance and metabolic dependencies. Additionally, this model supports studies on PPAR and AMPK signaling, mitochondrial dysfunction, and lipid metabolism reprogramming in HCC. For further details, please contact Ascent Research.
