The CBR1 Knockout HGC-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the human gastric carcinoma HGC-27 cell line, designed to disrupt the CBR1 gene encoding NADPH-dependent carbonyl reductase 1. This gene-edited cell pool provides a heterogenous knockout model for studying loss-of-function effects in a gastric cancer background, generated by CRISPR/Cas9-mediated targeted gene disruption without clonal isolation. The polyclonal format maintains genetic diversity while ensuring robust ablation of CBR1 expression, suitable for population-level functional analyses.
The HGC-27 parental cell line is an established human gastric adenocarcinoma line originally isolated from a lymph node metastasis of a gastric cancer patient. As gastric carcinoma epithelial cells, HGC-27 exhibits adherent growth and retains key characteristics of gastric cancer, including dysregulated proliferative signaling and altered metabolic pathways. This cellular context is particularly relevant for investigating CBR1 function in gastric tumor biology, where the enzyme contributes to drug metabolism and endogenous lipid mediator regulation.
CBR1 is an NADPH-dependent short-chain dehydrogenase/reductase that catalyzes the reduction of a broad spectrum of carbonyl-containing substrates, including quinones, prostaglandin E2 (PGE2), and diverse xenobiotics such as the chemotherapeutic agent doxorubicin. The enzyme functions downstream of transcriptional regulators NFE2L2 (Nrf2) and PPARG, and is responsive to TNF-mediated inflammatory signals. CBR1 interacts with NADPH as a cofactor and coordinates with phase I cytochrome P450 enzymes and phase II glutathione S-transferases in detoxification pathways. Within prostaglandin metabolism, CBR1 catalyzes the conversion of PGE2 to 15-keto-PGE2, thereby inactivating this pro-inflammatory mediator and modulating PTGS2 (COX-2)-driven signaling. In drug metabolism, CBR1 reduces doxorubicin to doxorubicinol, which is associated with cardiotoxic side effects and altered anticancer efficacy, and also participates in retinoid reduction impacting cellular differentiation and stress responses.
In the HGC-27 gastric cancer background, knockout of CBR1 disrupts these metabolic and signaling networks, providing a powerful tool to examine mechanisms of chemoresistance and oxidative stress. Loss of CBR1 may impair the inactivation of reactive carbonyl species and prostaglandins, potentially altering the cellular redox balance and inflammatory microenvironment. Given the role of CBR1 in doxorubicin metabolism, this model is especially valuable for dissecting the balance between drug efficacy and toxicity in gastric cancer. Furthermore, CBR1 knockout in this epithelial cell line allows investigation of prostaglandin-dependent signaling pathways critical for gastric mucosal homeostasis and tumor progression.
Researchers can employ these polyclonal knockout cells in a wide array of applications, including drug sensitivity assays (MTT/CCK8) to assess chemosensitivity to doxorubicin and other carbonyl-containing drugs, LC-MS-based quantification of prostaglandin metabolites to profile lipid mediator changes, and flow cytometric analysis of reactive oxygen species (ROS) to evaluate oxidative stress responses. Additional techniques such as Western blotting and RT-qPCR enable confirmation of CBR1 ablation and its downstream targets, while transcriptomic analyses (RNA-seq) can uncover global pathway alterations. Migration assays may also be conducted to study the impact on invasive potential. These applications support investigations into gastric cancer drug resistance, xenobiotic metabolism, and redox signaling. For additional details, please contact Ascent Research.
