The AACS Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 hepatocellular carcinoma cell line, featuring targeted disruption of the AACS gene. This product comprises a heterogeneous pool of cells with CRISPR/Cas9-mediated gene disruption, enabling loss-of-function studies without clonal isolation. The population maintains parental Huh-7 properties while lacking AACS protein expression, as verified by Western blot.
Huh-7 cells originate from a liver tumor of a 57-year-old Japanese male and serve as a well-established model for hepatocellular carcinoma. They retain features such as unregulated proliferation, altered metabolism, and drug sensitivity, making them ideal for investigating liver cancer biology and therapeutic responses. This context is particularly relevant for exploring metabolic gene functions in HCC.
AACS encodes acetoacetyl-CoA synthetase, which catalyzes the conversion of acetoacetate to acetoacetyl-CoA, linking ketone body catabolism to lipid synthesis and cholesterol biosynthesis. Regulated by PPAR??, insulin, glucagon, and SREBP-1c, AACS functions downstream of dietary ketones and utilizes acetoacetate, coenzyme A, and ATP. The product acetoacetyl-CoA enters the mevalonate pathway via HMGCS2 to generate HMG-CoA, mevalonate, isoprenoids, and cholesterol, as well as contributing to fatty acid synthesis. Thus, AACS sits at the intersection of ketone body utilization and anabolic lipid metabolism.
In Huh-7 cells, AACS knockout disrupts ketone body re-utilization for lipid and cholesterol production, impairing membrane biosynthesis and potentially reducing cancer cell proliferation under metabolic stress. This model illuminates how HCC cells exploit ketone bodies for anabolic demands and may reveal metabolic dependencies for therapeutic targeting, especially concerning the mevalonate pathway.
Key applications include metabolic reprogramming in HCC, ketone body utilization, and cholesterol biosynthesis. Assays such as Western blot, RT-qPCR, cholesterol measurement, fatty acid oxidation, cell proliferation, Oil Red O staining, Seahorse mitochondrial respiration, LC-MS metabolomics, and statin sensitivity testing can be employed. For further information, please contact Ascent Research.
