The CBR3 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human hepatic adenocarcinoma cell line. This polyclonal knockout model is engineered for targeted disruption of the CBR3 gene, which encodes NADPH-dependent carbonyl reductase 3, an enzyme that reduces a broad spectrum of carbonyl-containing substrates. The heterogeneous edited population provides a robust system for studying CBR3-dependent processes without the clonal bias inherent in single-cell-derived lines, making it ideal for functional genomics, pathway analysis, and drug discovery applications.
The SK-HEP-1 host cell line, derived from a human liver adenocarcinoma, is characterized by an endothelial-like phenotype and displays markers of liver sinusoidal endothelial cells (LSECs), making it a pertinent model for liver cancer research and endothelial biology. The cells maintain hepatic lineage features alongside endothelial characteristics, enabling studies of tumor microenvironment interactions and metabolic pathways that bridge oncogenic and vascular functions. This background offers a unique platform for investigating how carbonyl metabolism intersects with liver sinusoidal endothelial cell biology and hepatocellular carcinoma progression.
CBR3 functions as a monomeric carbonyl reductase that utilizes NADPH to reduce prostaglandins, quinones, and xenobiotic aldehydes, contributing to cellular detoxification and oxidative stress responses. Its expression is transcriptionally regulated by the Nrf2?CKEAP1 antioxidant pathway and the aryl hydrocarbon receptor (AhR), both activated by reactive oxygen species (ROS). In the SK-HEP-1 setting, CBR3 operates within a network that includes prostaglandin-endoperoxide synthase 2 (PTGS2/COX-2), prostaglandin E synthase (PTGES), 15-hydroxyprostaglandin dehydrogenase (HPGD), and NAD(P)H:quinone oxidoreductase 1 (NQO1). Disruption of CBR3 impairs the reduction of 15-keto-prostaglandin E2 (15-keto-PGE2) and other carbonyl metabolites, leading to accumulation of reactive carbonyl species, altered prostaglandin metabolism, and dysregulated oxidative stress signaling that influences inflammatory pathways.
In SK-HEP-1 cells, CBR3 knockout enables dissection of carbonyl metabolism in the context of liver cancer and endothelial biology, potentially impacting drug transport and metabolic zonation. The loss of CBR3-mediated detoxification can sensitize these cells to chemotherapeutic agents and electrophilic stress, offering a model for investigating drug resistance in hepatocarcinoma, breast, and lung cancers. Moreover, the dual hepatic?Cendothelial nature allows examination of carbonyl reduction in angiogenic signaling and tumor?Cstromal interactions, potentially uncovering new therapeutic targets.
The polyclonal knockout cells support diverse assays, including Western blotting and RT-qPCR for CBR3 verification, carbonyl reductase activity measurements, LC-MS-based prostaglandin metabolite profiling, ROS detection with H2DCFDA, and MTT drug sensitivity testing. They are also suitable for immunofluorescence to monitor Nrf2 nuclear translocation and for RNA-seq transcriptomic analysis. Key research applications encompass carbonyl metabolism, oxidative stress, prostaglandin signaling, cancer metabolism, and toxicology screening. For further technical details or ordering information, please contact Ascent Research.
