The HIBADH Knouckout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting HIBADH in the SK-HEP-1 human hepatic adenocarcinoma cell line. This heterogeneous pool contains cells with gene disruption at the HIBADH locus, enabling loss-of-function studies without single-cell cloning. It provides a robust model for investigating branched-chain amino acid metabolism in a liver cancer background.
SK-HEP-1 cells, derived from the ascites of a hepatic adenocarcinoma patient, are widely used as a liver cancer model. This line exhibits metabolic features of hepatic tumors, including altered amino acid utilization and mitochondrial activity, making it suitable for dissecting valine catabolism??s role in cancer metabolism. Its hepatic origin offers a disease-relevant context for studying how HIBADH disruption affects metabolic flux and energy homeostasis.
HIBADH encodes mitochondrial 3-hydroxyisobutyrate dehydrogenase, catalyzing the NAD-dependent oxidation of 3-hydroxyisobutyrate to methylmalonate semialdehyde in the valine degradation pathway, thereby linking branched-chain amino acid breakdown to the TCA cycle and NADH-dependent respiration. The homodimeric enzyme interacts with downstream ALDH6A1 to process methylmalonate semialdehyde to propionyl-CoA. Activity is modulated by substrate availability, NAD+/NADH ratio, and transcriptional regulators like PPARGC1A. HIBADH disruption causes 3-hydroxyisobutyrate accumulation, reduced TCA cycle flux, and potential mitochondrial dysfunction.
In SK-HEP-1 hepatic adenocarcinoma cells, HIBADH knockout permits study of branched-chain amino acid metabolism??s support of tumor proliferation and survival. Aberrant valine catabolism is implicated in cancer metabolic reprogramming; HIBADH loss may sensitize cells to nutrient deprivation or alter redox. The model is valuable for investigating 3-hydroxyisobutyric aciduria, organic acidurias, and links to metabolic syndrome, allowing delineation of HIBADH??s contribution to liver cancer metabolism and identification of therapeutic targets.
The polyclonal HIBADH knockout SK-HEP-1 cells are suited for diverse applications. Enzyme activity assays and LC-MS quantification of 3-hydroxyisobutyrate confirm functional knockout, while RT-qPCR, western blotting, and Seahorse mitochondrial stress tests assess expression and bioenergetic changes. Proliferation, viability, and apoptosis assays under nutrient limitation reveal metabolic dependencies. This model supports metabolic flux analysis, drug target validation, and mechanistic studies. For further details, please contact Ascent Research.
