The H6PD Knockout NCI-H1703 Polyclonal Cells consist of a polyclonal population of the NCI-H1703 human non-small cell lung cancer line, engineered via CRISPR/Cas9 to disrupt the H6PD gene. These polyclonal knockout cells provide a loss-of-function model that circumvents clonal selection, enabling studies of H6PD-dependent biological processes in a heterogeneous cell background. The CRISPR-mediated gene inactivation targets H6PD, which encodes hexose-6-phosphate dehydrogenase, an enzyme critical for generating NADPH within the endoplasmic reticulum.
NCI-H1703 cells originate from a pleural effusion metastasis of a male patient with lung squamous cell carcinoma. This epithelial cell line harbors well-characterized oncogenic mutations in KRAS and TP53, making it a clinically relevant in vitro model for non-small cell lung cancer biology, particularly for investigating signaling pathways and drug responses. These cells are commonly used to study tumor cell survival mechanisms and metabolic adaptations, providing a suitable platform for examining how loss of H6PD influences lung cancer pathophysiology.
H6PD functions in the endoplasmic reticulum lumen, oxidizing glucose-6-phosphate to produce NADPH. The elevated NADPH/NADP+ ratio allosterically activates 11??-hydroxysteroid dehydrogenase type 1 (HSD11B1), which reduces cortisone to active cortisol, amplifying glucocorticoid receptor signaling. The pathway is regulated by upstream inputs including oxidative stress, glucocorticoids, and the unfolded protein response mediators ATF6 and XBP1. Downstream, H6PD controls the NADPH pool, HSD11B1 reductase activity, and redox-sensitive ER chaperones. Key interacting factors include HSD11B1, the glucose-6-phosphate transporter, and NADP+, integrating hexose-6-phosphate metabolism with ER redox homeostasis and local cortisol production.
In KRAS/TP53-mutant NCI-H1703 cells, H6PD-driven NADPH generation and cortisol synthesis may support proliferation, survival under oxidative stress, and metabolic adaptation. This knockout model is particularly valuable for dissecting the interplay between ER redox regulation and oncogenic signaling, clarifying how H6PD influences tumor cell fitness, drug sensitivity, and the broader stress response in non-small cell lung cancer.
Researchers can employ this product in a variety of assays, including western blotting and RT-qPCR to confirm H6PD ablation, NADPH/NADP+ ratio assays, LC-MS/MS-based cortisol/cortisone quantification, and HSD11B1 reductase activity measurements. Functional analyses such as cell proliferation, oxidative stress response, apoptosis, and clonogenic survival studies enable thorough assessment of H6PD??s role in lung cancer cell resilience. Moreover, the model facilitates exploration of H6PD as a potential therapeutic target and its involvement in drug resistance mechanisms. For further information, please contact Ascent Research.
