This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human hepatocellular carcinoma cell line, in which the HYI gene has been disrupted to eliminate functional hydroxypyruvate isomerase expression. The polyclonal format provides a heterogeneous knockout pool that better recapitulates population-level metabolic responses compared to a single clone, making it suitable for investigating the role of HYI in glyoxylate metabolism without the confounding effects of clonal selection. Researchers can use this model to examine the functional consequences of HYI loss in an epithelial liver adenocarcinoma background.
The SK-HEP-1 cell line was originally established from the ascitic fluid of a patient with liver adenocarcinoma and exhibits an epithelial morphology. It is characterized by wild-type KRAS status and is frequently employed in hepatocellular carcinoma (HCC) research, as well as in studies of hepatic metabolic function. The cells retain many features of liver parenchymal cells and are responsive to metabolic stimuli, providing a physiologically relevant context for investigating the impact of HYI disruption on cancer metabolism and glyoxylate detoxification pathways. Importantly, SK-HEP-1 cells are amenable to a wide range of molecular and metabolic assays, enabling robust functional characterization.
The HYI gene encodes a mitochondrial hydroxypyruvate isomerase that catalyzes the reversible isomerization of hydroxypyruvate to 2-hydroxy-3-oxopropanoate. This enzyme functions downstream of AGXT and GRHPR in the glyoxylate and dicarboxylate metabolism pathway, and its activity is putatively regulated by metabolic transcription factors such as PPAR?? and HNF4A. Disruption of HYI is predicted to impair glyoxylate detoxification, leading to accumulation of toxic aldehydes and perturbation of mitochondrial redox balance. The enzyme complexes with its substrate hydroxypyruvate and product 2-hydroxy-3-oxopropanoate, and cooperates with HOGA1 to prevent oxalate overproduction, highlighting its critical role in maintaining metabolic homeostasis.
In the context of SK-HEP-1 hepatocellular carcinoma cells, knockout of HYI provides a powerful tool to dissect the role of glyoxylate metabolism in liver cancer biology. HCC cells frequently rewire their metabolic pathways to sustain proliferation and survive under stress; loss of HYI-dependent glyoxylate detoxification may render these cells vulnerable to mitochondrial dysfunction and oxidative damage. This polyclonal model enables researchers to evaluate how glyoxylate accumulation influences tumor cell viability, redox balance, and metabolic flux, linking HYI function to oncogenic metabolic adaptation. Moreover, it serves as a relevant hepatic platform for modeling hyperoxaluria-related cellular pathology, as SK-HEP-1 cells retain metabolic competencies required for oxalate precursor handling.
Typical applications include western blot, RT-qPCR, and hydroxypyruvate isomerase activity assays for knockout validation, as well as glyoxylate quantification and metabolic flux analysis using isotope-labeled substrates. Functional studies can assess mitochondrial respiration via Seahorse and cell viability under glyoxylate-induced stress, while combination with pharmacological inhibitors enables evaluation of HYI as a therapeutic target in hyperoxaluria and HCC. For further information, please contact Ascent Research.
