The H6PD Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the SK-HEP-1 human hepatocellular carcinoma line. This product features CRISPR/Cas9-mediated disruption of the H6PD gene, yielding a heterogeneous pool of cells with loss-of-function mutations. Unlike monoclonal lines, this polyclonal format maintains population-level heterogeneity while eliminating H6PD activity, providing a robust tool for pathway studies. CRISPR/Cas9 targeting ensures precise gene disruption.
The SK-HEP-1 cell line was established from ascitic fluid of a liver adenocarcinoma patient and displays epithelial morphology. As a hepatocellular carcinoma model, it retains features such as dysregulated metabolism and altered signaling, and is widely used to study tumor progression and therapeutic response. Its hepatic origin makes it suitable for examining pathways like glucocorticoid metabolism and ER redox processes.
H6PD encodes an ER-luminal enzyme that converts glucose-6-phosphate to generate NADPH in the pentose phosphate pathway. This NADPH fuels HSD11B1, which reduces cortisone to cortisol, thereby activating glucocorticoid receptor (GR) signaling. H6PD expression is regulated by insulin, PPAR?? agonists, and glucose, and controls downstream outputs including HSD11B1 activity, cortisol production, and the NADPH/NADP+ ratio. Interacting factors include NADP+ and hexose-6-phosphate. Thus, H6PD serves as a crucial metabolic gatekeeper for corticosteroid metabolism.
In SK-HEP-1 liver cancer cells, H6PD knockout disrupts cortisol generation and GR activation, potentially affecting proliferation, redox balance, and metabolic reprogramming. Glucocorticoid signaling is implicated in obesity-associated cancers and metabolic syndrome; by ablating H6PD, this model enables investigation of how ER NADPH and cortisol influence hepatocellular carcinoma phenotypes. The altered NADPH/NADP+ ratio may also impact drug resistance and redox homeostasis.
Applications include studying glucocorticoid metabolism in HCC, metabolic reprogramming, and redox biology. Standard assays like RT-qPCR, Western blotting, cortisol ELISA, NADPH/NADP+ measurements, MTT, ROS detection, and RNA-seq can be employed. The polyclonal format supports population-level analyses of gene expression, chromatin binding (ChIP-qPCR), and protein interactions (co-immunoprecipitation). Glucose uptake assays further characterize metabolic shifts. For additional details, contact Ascent Research.
