The GPX1 Knockout SK-HEP-1 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population derived from the SK-HEP-1 cell line, featuring targeted disruption of the GPX1 gene. This heterogeneous pool of edited cells provides a relevant loss-of-function model for studying glutathione peroxidase 1 function without the need for single-cell cloning, preserving biological variability inherent to the parental line. The knockout is achieved through CRISPR/Cas9-mediated gene disruption, enabling robust investigation of GPX1-dependent processes in a liver adenocarcinoma background.
The host SK-HEP-1 cell line was established from the ascitic fluid of a patient with liver adenocarcinoma and displays an endothelial-like phenotype. This hepatic adenocarcinoma cell model is widely employed in hepatocellular carcinoma research and studies of endothelial biology. Its origin from metastatic ascites makes it particularly suitable for exploring tumor cell survival mechanisms under stress conditions, including oxidative and nutrient challenges relevant to the tumor microenvironment.
GPX1 encodes an essential antioxidant enzyme that catalyzes the reduction of hydrogen peroxide and organic hydroperoxides to water or corresponding alcohols, using glutathione as a cofactor. GPX1 is transcriptionally regulated by NFE2L2/NRF2 and TP53, and its activity depends on selenium availability and the AP-1 complex. It functions within the glutathione metabolism and antioxidant defense pathways, directly contributing to redox homeostasis. The enzyme interacts with superoxide dismutase 1 (SOD1), catalase, glutathione reductase, and thioredoxin to coordinate reactive oxygen species (ROS) detoxification. Downstream, GPX1 modulates signaling through the PI3K/AKT, MAPK/ERK, and NF-??B pathways, and indirectly influences BCL2 family proteins, thereby linking oxidative stress to cell survival and apoptotic decisions.
In the SK-HEP-1 hepatic adenocarcinoma context, GPX1 disruption leads to accumulation of hydrogen peroxide and lipid peroxides, sensitizing cells to oxidative stress and ferroptosis. This redox imbalance may impair tumor cell survival and promote apoptosis under pro-oxidant conditions, making the model valuable for dissecting ferroptotic death mechanisms and assessing therapeutic vulnerabilities in liver cancer. The polyclonal knockout pool captures heterogeneous editing outcomes, reflecting a realistic range of functional perturbations and overcoming clonal selection biases often encountered in isogenic lines.
These polyclonal knockout cells are suited for diverse experimental applications, including investigation of oxidative stress responses, ferroptosis induction studies with agents such as erastin or RSL3, and drug sensitivity assays for pro-oxidant therapies targeting hepatocellular carcinoma. Users can assess GPX activity, measure ROS levels via DCFH-DA, quantify lipid peroxidation using MDA assays, or perform western blotting to monitor downstream signaling changes. The model also supports cell viability assays under hydrogen peroxide challenge, enabling detailed dissection of redox-controlled pathways in liver adenocarcioma. For further technical information and ordering details, please contact Ascent Research.
