The GSR knockout SK-HEP-1 polyclonal cells are a CRISPR/Cas9-edited polyclonal population derived from the SK-HEP-1 human hepatic adenocarcinoma cell line, designed for functional studies of glutathione reductase (GSR). This heterogeneous pool contains various GSR-disrupted alleles, providing a robust loss-of-function model without clonal bias. The polyclonal format is ideal for investigating redox biology in a cancer context and for applications requiring bulk population-level responses.
SK-HEP-1 is a human hepatic adenocarcinoma cell line originally derived from ascites fluid of a patient with hepatocellular carcinoma. It displays epithelial morphology and is extensively used as a model for liver cancer, including studies of tumor progression, metastasis, and drug resistance. The line retains characteristic oncogenic signaling and metabolic features of hepatic malignancies, making it an appropriate host for investigating the role of glutathione metabolism in cancer redox adaptation.
GSR encodes glutathione reductase, which catalyzes NADPH-dependent reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH), maintaining the GSH/GSSG ratio essential for redox homeostasis. GSR is transcriptionally regulated by NFE2L2 (NRF2), AP-1, HIF1A, and PPARGC1A, linking antioxidant responses to cellular stress and metabolism. The enzyme operates in concert with NADPH-generating pathways and interacts with GSH, glutaredoxin, and thioredoxin systems. Downstream, sustained GSH pools support peroxiredoxins, glutathione S-transferases, and S-glutathionylation reactions, collectively controlling detoxification and redox signaling.
Disruption of GSR in SK-HEP-1 cells causes GSSG accumulation and GSH depletion, compromising ROS scavenging and shifting the redox balance towards a pro-oxidant state. This sensitizes the cells to apoptosis and ferroptosis, particularly under exogenous oxidative stress or ferroptosis inducers like erastin. In the hepatocellular carcinoma context, this knockout model enables dissection of redox-dependent survival mechanisms and the role of glutathione recycling in cancer cell resilience. It also facilitates exploration of how redox dysregulation intersects with oncogenic signaling to uncover therapeutic targets.
This polyclonal GSR knockout cell population is suitable for a comprehensive array of functional assays, including measurement of GSH/GSSG ratios, detection of intracellular ROS with fluorescent probes, and cell viability dose-response curves under oxidative stress or ferroptosis induction by compounds such as erastin. It can be employed in drug screening to identify molecules that selectively target GSR-deficient cancer cells, and in transcriptomic (RNA-seq) or proteomic analyses to map adaptive redox responses. Standard validation by Western blotting and RT-qPCR confirms GSR disruption and reveals potential upregulation of compensatory pathways. For detailed protocols, pricing, or collaborative inquiries, please contact Ascent Research.
