The H6PD Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-mediated gene-disrupted polyclonal population designed to abolish H6PD expression in the human NCI-H1975 lung adenocarcinoma cell line. This product provides a powerful loss-of-function model to interrogate the roles of hexose-6-phosphate dehydrogenase in endoplasmic reticulum (ER) luminal NADPH generation and downstream glucocorticoid metabolism. As a polyclonal knockout pool, it circumvents the clonal variability often encountered with single-cell-derived knockouts, offering a more representative population-level assessment of H6PD-dependent phenotypes. Researchers can utilize these cells to dissect the intersection of the pentose phosphate pathway with redox homeostasis and hormone activation.
NCI-H1975 is a widely utilized non-small cell lung cancer (NSCLC) line derived from the metastatic pleural effusion of a female nonsmoker. It harbors activating EGFR mutations (L858R/T790M), which confer both oncogene addiction and resistance to first-generation tyrosine kinase inhibitors, making it a standard model for studying EGFR-targeted therapy and acquired resistance. The epithelial origin and tumorigenic properties of NCI-H1975 provide a clinically relevant context for investigating metabolic and signaling adaptations in lung adenocarcinoma. Its well-characterized molecular background ensures reproducible experimental outcomes.
H6PD is an ER-resident enzyme that catalyzes the oxidation of glucose-6-phosphate (G6P) to 6-phosphogluconolactone (6PGL), generating NADPH within the ER lumen. This NADPH pool is essential for the reductase activity of 11??-hydroxysteroid dehydrogenase type 1 (11??-HSD1/HSD11B1), which converts inert cortisone to active cortisol. Thus, H6PD functions upstream of 11??-HSD1 as an obligate redox partner, and their interaction enables tissue-specific glucocorticoid amplification. Transcriptional regulation by glucocorticoids and ER stress links H6PD to cellular energy status. Downstream, cortisol activates glucocorticoid receptors to modulate genes involved in metabolism and inflammation. Disruption of H6PD depletes luminal NADPH, impairing 11??-HSD1 activity and altering local glucocorticoid balance.
Ablating H6PD in the NCI-H1975 background offers unique insights into how local glucocorticoid activation intersects with oncogenic signaling. Lung adenocarcinoma cells often face fluctuating redox and nutrient stress, and H6PD-dependent NADPH production may buffer oxidative challenges. By knocking out H6PD, researchers can delineate its contribution to maintaining redox balance and supporting proliferation under metabolic constraints. Moreover, since NCI-H1975 cells express EGFR mutants that drive downstream cascades, the model enables exploration of crosstalk between growth factor signaling and glucocorticoid metabolism. This targeted knockout may reveal vulnerabilities in NSCLC that could inform combination therapeutic strategies.
The H6PD Knockout NCI-H1975 Polyclonal Cells are suited for diverse investigations. Key applications include glucocorticoid metabolism studies via cortisol/cortisone LC-MS and 11??-HSD1 enzyme activity assays. For metabolic research, this model enables NADPH/NADP+ ratio measurement and analysis of redox-dependent processes like ROS defense and lipogenesis. Cancer biologists can examine ER redox impacts on tumor fitness and drug resistance using western blotting and RT-qPCR. Immunofluorescence validates H6PD and 11??-HSD1 localization. Overall, this polyclonal knockout is a robust tool for exploring ER redox biology and glucocorticoid action in lung cancer. Contact Ascent Research for more information.
