The H6PD Knockout UM-UC-3 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the H6PD gene in the human UM-UC-3 cell line. This polyclonal knockout model provides a heterogeneous pool of cells, each carrying targeted gene disruptions introduced by CRISPR/Cas9 technology, enabling functional loss-of-function studies without the need for clonal isolation. The product is suitable for experiments requiring a population-level knockout effect, offering a versatile tool for investigating the biological role of H6PD in cancer biology and redox metabolism.
The parental UM-UC-3 cell line was established from a male patient with metastatic bladder transitional cell carcinoma and serves as a widely used in vitro model for bladder cancer research. These epithelial cells retain key characteristics of the original tumor, making them valuable for studying tumorigenesis, metastasis, and drug responses. The availability of H6PD knockout in this cellular context allows researchers to dissect how H6PD-mediated NADPH production influences the malignant phenotype of bladder cancer cells.
H6PD (hexose-6-phosphate dehydrogenase) resides in the endoplasmic reticulum (ER) lumen and catalyzes the oxidation of glucose-6-phosphate to generate NADPH, a critical cofactor for ER redox homeostasis and reductive biosynthesis. Mechanistically, H6PD-derived NADPH drives the oxoreductase activity of 11??-hydroxysteroid dehydrogenase type 1 (HSD11B1), which converts inert cortisone to active cortisol. This local cortisol regeneration amplifies glucocorticoid receptor (GR) signaling independently of circulating corticosteroid levels. H6PD is transcriptionally regulated by glucocorticoids themselves, SREBP-1c, and PPAR??, forming a feed-forward loop that sustains glucocorticoid action. Key downstream effects include modulation of HSD11B1 activity, local GR signaling, and maintenance of ER redox balance. Interacting partners include HSD11B1 and various ER oxidoreductases, highlighting the integration of H6PD into corticosteroid metabolism and NADP/NADPH homeostasis.
In the context of UM-UC-3 bladder cancer cells, H6PD is positioned at the nexus of NADPH metabolism and glucocorticoid signaling, two pathways implicated in tumor progression and therapeutic resistance. Disruption of H6PD can perturb the local generation of cortisol, potentially attenuating GR-mediated transcriptional programs that promote proliferation, survival, and metabolic adaptation. Moreover, impaired ER NADPH production may sensitize cancer cells to oxidative stress and genotoxic agents, offering a model to elucidate the role of redox regulation in bladder cancer. This knockout model thus enables the study of H6PD-dependent mechanisms that contribute to the aggressive phenotype of bladder transitional cell carcinoma.
The H6PD Knockout UM-UC-3 Polyclonal Cells are suited for a broad range of functional assays. Researchers can quantify intracellular NADPH levels using enzymatic cycling or fluorescence-based assays, and monitor cortisol-to-cortisone ratios by LC-MS to assess HSD11B1 reductase activity. Western blotting and immunofluorescence detect H6PD and HSD11B1 protein expression, while RT-qPCR or RNA-seq can profile glucocorticoid-responsive gene signatures. Cellular proliferation and apoptosis assays help evaluate the impact of H6PD loss on tumor cell fitness, and the model is applicable to drug screening for cortisone reductase deficiency or related metabolic disorders. For additional information or customized solutions, please contact Ascent Research.
