The H6PD Knockout LoVo Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population designed for targeted disruption of the hexose-6-phosphate dehydrogenase (H6PD) gene in the human LoVo colorectal adenocarcinoma line. This loss-of-function model abrogates H6PD-dependent glucose-6-phosphate dehydrogenase activity in the endoplasmic reticulum (ER), enabling precise analysis of NADPH generation, glucocorticoid activation, and redox homeostasis. As a heterogeneous polyclonal pool, the knockout preserves LoVo epithelial features while circumventing clonal selection artifacts.
The LoVo cell line, established from a metastatic colorectal adenocarcinoma of a 56-year-old male, is a well-characterized epithelial model for colorectal cancer. Exhibiting characteristic mutations in oncogenes and tumor suppressors, LoVo cells are widely applied in studies of tumor cell signaling, metastatic behavior, and metabolic rewiring. Their colonic origin provides a physiologically relevant context for investigating how local glucocorticoid metabolism impacts intestinal tumor biology.
H6PD encodes a bifunctional ER enzyme that imports glucose-6-phosphate and oxidizes it to 6-phosphogluconolactone, generating NADPH. This luminal NADPH is an essential cofactor for 11??-hydroxysteroid dehydrogenase type 1 (HSD11B1), which converts inactive cortisone to active cortisol. Upstream, H6PD expression is regulated by PPAR??, C/EBP??, ER stress, and substrate availability. Downstream, H6PD activity modulates the NADPH/NADP+ ratio, cortisol production, and the glucose-6-phosphate pool, positioning the H6PD-HSD11B1 axis as a key regulator of local glucocorticoid activation.
In LoVo colorectal cancer cells, disruption of H6PD eliminates the ER-driven supply of NADPH for cortisol synthesis, offering a strategic tool to dissect the contribution of local glucocorticoid production to tumor cell proliferation, survival, and drug sensitivity. Because H6PD and glucocorticoid dysregulation are implicated in metabolic syndrome, obesity, and type 2 diabetes??conditions that elevate colorectal cancer risk??this knockout model facilitates research into the molecular crosstalk between metabolic disorders and tumor progression.
The polyclonal knockout cells support a range of analytical approaches, including LC-MS measurement of cortisol/cortisone ratios, H6PD activity assays, NADPH quantification, western blotting for HSD11B1/H6PD, RT-qPCR, and glucose-6-phosphate uptake studies. Applications span glucocorticoid metabolism research, HSD11B1 inhibitor screening, cortisone reductase deficiency modeling, and ER redox biology. For further information or to discuss customization, please contact Ascent Research.
