The H6PD Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population derived from the HCT 116 human colorectal carcinoma cell line, engineered to disrupt the H6PD gene which encodes hexose-6-phosphate dehydrogenase. This polyclonal product enables loss-of-function studies without single-cell cloning, providing a heterogeneous genetic background that better reflects population-level responses in cancer biology. The knockout model is designed for investigating the molecular consequences of impaired ER NADPH generation on glucocorticoid metabolism and redox homeostasis.
The parental HCT 116 cell line is a widely utilized model of colorectal carcinoma, originally derived from an adult male patient. These cells harbor a KRAS G13D gain-of-function mutation and a CTNNB1??S45 deletion that stabilizes ??-catenin, resulting in constitutive Wnt pathway activation. HCT 116 cells are near-diploid, facilitating efficient gene targeting, and their defined oncogenic landscape makes them particularly suitable for dissecting signaling cross-talk and metabolic adaptations in colorectal cancer.
Mechanistically, H6PD catalyzes the oxidation of glucose-6-phosphate to 6-phosphogluconolactone within the endoplasmic reticulum lumen, using NADP+ as a cofactor to generate NADPH. This luminal NADPH pool is indispensable for the reductase activity of HSD11B1, which converts inactive cortisone to active cortisol, thereby regulating glucocorticoid signaling. H6PD activity is transcriptionally regulated by NRF2 and PPAR?? in response to oxidative stress and functionally cooperates with the G6P transporter SLC37A4 (G6PT) to ensure substrate availability. Downstream, NADPH produced by H6PD supports ER redox-sensitive proteins and various NADPH-dependent enzymes, positioning H6PD at a critical junction between energy metabolism, redox control, and glucocorticoid activation.
In the HCT 116 colorectal carcinoma context, co-occurring KRAS and ??-catenin mutations impose heightened metabolic and oxidative demands on the ER. Disruption of H6PD depletes the ER NADPH pool, impairing HSD11B1-mediated cortisol production and compromising the cellular ability to maintain ER redox balance. Consequently, this knockout model is highly relevant for elucidating the interplay between oncogenic signaling, ER redox homeostasis, and glucocorticoid metabolism. It may uncover adaptive mechanisms to oxidative stress and expose metabolic vulnerabilities exploitable for therapeutic intervention in colorectal cancer and metabolic syndrome.
Researchers can employ this polyclonal knockout population to investigate glucocorticoid-driven survival pathways in colorectal cancer by quantifying cortisol production via LC-MS and monitoring HSD11B1 expression through western blot or RT-qPCR. The model is well suited for assessing ER redox dynamics using NADPH/NADP+ ratio measurements and for evaluating sensitivity to oxidative stress inducers through cell viability and flow cytometry-based apoptosis assays. Furthermore, colony formation assays can be applied to examine clonogenic survival under metabolic or pharmacological stress. For further technical details and product inquiries, please contact Ascent Research.
