The IDH3B Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from human SK-HEP-1 hepatocellular carcinoma cells. This heterogeneous pool comprises cells with distinct loss-of-function mutations in IDH3B, generated by CRISPR/Cas9-mediated gene disruption. The polyclonal format minimizes clonal bias and provides a robust model for studying IDH3B functional ablation. These cells are suited for investigating immediate and population-level metabolic effects.
SK-HEP-1 is a human hepatocellular carcinoma cell line originally derived from the ascitic fluid of a 52-year-old male with liver adenocarcinoma. The cells display adherent epithelial morphology and retain key metabolic and signaling features relevant to hepatic tumor biology. Their mesenchymal traits also facilitate studies on cancer cell plasticity. This background renders the model appropriate for probing metabolic dysregulation in liver cancer.
IDH3B encodes the beta subunit of mitochondrial NAD+-dependent isocitrate dehydrogenase (IDH3), a TCA cycle enzyme that converts isocitrate to ??-ketoglutarate (??-KG) with concomitant NADH production. The IDH3 complex assembles from IDH3A, IDH3B, and IDH3G subunits. Transcription factors MYC, HIF1A, and PPARGC1A regulate IDH3B expression. IDH3 acts downstream of aconitase 2 (ACO2) and upstream of ??-ketoglutarate dehydrogenase (OGDH). ??-KG functions as a co-substrate for dioxygenases involved in DNA and histone demethylation, connecting mitochondrial metabolism to epigenetics. NADH generated by IDH3 feeds complex I of the electron transport chain. IDH3B disruption therefore diminishes ??-KG and NADH output, impairing biosynthetic pathways and redox homeostasis.
In SK-HEP-1 hepatocellular carcinoma cells, IDH3B knockout offers a model to dissect metabolic reprogramming in liver cancer. Hepatocellular carcinomas often rewire the TCA cycle to support growth. IDH3B deficiency may force reliance on glutaminolysis or reductive carboxylation, revealing metabolic vulnerabilities. This model can also be used to examine interactions between altered TCA cycle activity and common liver cancer mutations.
Typical applications include confirmation of knockout by Western blot or RT-qPCR, metabolite profiling by LC-MS, and Seahorse respirometry to quantify mitochondrial function. NADH/NAD+ ratio assays and proliferation/colony formation analyses further elucidate the functional consequences. These polyclonal knockout cells are thus a versatile tool for investigating TCA cycle dynamics, metabolic adaptations in hepatocellular carcinoma, and the epigenetic and bioenergetic impacts of IDH3B loss. For further technical inquiries, please contact Ascent Research.
