The H6PD Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the CAL-27 human tongue squamous cell carcinoma line. This product features targeted disruption of the H6PD gene, which encodes hexose-6-phosphate dehydrogenase, without selection for clonal homogeneity. The heterogeneous pool preserves the host cell background while introducing loss-of-function mutations at the target locus, providing a ready-to-use model for population-level studies of H6PD deficiency.
The host cell line CAL-27 originates from a human male oral squamous cell carcinoma of the tongue, serving as a widely used preclinical model for head and neck cancer research. These adherent epithelial cells harbor common oral cancer mutations and exhibit robust in vitro growth, making them suitable for functional assays. The knockout is performed in this clinically relevant background, ensuring phenotype attribution to H6PD loss.
H6PD encodes an ER-luminal enzyme that catalyzes glucose-6-phosphate conversion to 6-phosphogluconolactone, generating NADPH essential for ER redox homeostasis and steroid metabolism. This NADPH pool drives the reductase activity of 11??-hydroxysteroid dehydrogenase type 1 (HSD11B1), enabling cortisone-to-cortisol conversion, and supports ER oxidoreductases and chaperones such as GRP78 and calnexin. H6PD activity requires the glucose-6-phosphate transporter SLC37A4. Upstream regulation involves unfolded protein response mediators (PERK, IRE1), glucocorticoid receptor (NR3C1), and mTOR signaling. Downstream, NADPH availability influences cortisol production and multiple NADPH-dependent enzymes, positioning H6PD as a nexus between glucose metabolism, ER stress adaptation, and glucocorticoid signaling.
In CAL-27 oral cancer cells, H6PD disruption compromises ER luminal NADPH generation, impairing HSD11B1-mediated cortisol biosynthesis and sensitizing cells to oxidative stress. This deficiency may alter redox-dependent survival mechanisms, as tongue squamous cell carcinoma cells exploit antioxidant pathways to withstand metabolic and therapeutic challenges. The polyclonal knockout model enables investigation of H6PD-dependent glucocorticoid metabolism and ER redox control in cancer cell resilience, drug resistance, and apoptosis. Phenotypic assessments may include altered proliferation, increased ROS, and modulated stress signaling.
Applications include studying oral cancer metabolism, ER stress, and glucocorticoid signaling through NADPH/NADP+ ratio assays, cortisol ELISA, cell viability (MTT), ROS detection (DCFDA), and immunoblotting or immunofluorescence for ER markers. The model also facilitates H6PD inhibitor screening and exploration of redox vulnerabilities in squamous carcinoma. By combining H6PD perturbation with the CAL-27 background, researchers can link ER glucose metabolism to tumor-promoting pathways. For further details, contact Ascent Research.
