H6PD Knockout T-47D Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal cell population featuring targeted disruption of the H6PD gene in the human T-47D breast ductal carcinoma epithelial cell line. This knockout model is generated using CRISPR/Cas9-mediated gene disruption, resulting in a heterogeneous pool of cells carrying loss-of-function mutations in the H6PD locus. The polyclonal format preserves genetic diversity while ensuring functional ablation of hexose-6-phosphate dehydrogenase, offering a robust tool for studying glucocorticoid metabolism in hormone-responsive breast cancer without the clonal selection biases associated with monoclonal lines.
The parental T-47D cell line, derived from a pleural effusion of a human breast ductal carcinoma, is a well-characterized estrogen receptor-positive (ER+) and progesterone receptor-positive (PR+) luminal A breast cancer model. T-47D cells are widely employed for investigating hormone-dependent proliferation, steroid receptor signaling, and endocrine therapy response, making them an ideal chassis for dissecting the interplay between glucocorticoid and estrogen pathways in the luminal A molecular subtype.
H6PD encodes hexose-6-phosphate dehydrogenase, an endoplasmic reticulum (ER) luminal enzyme that generates NADPH via the pentose phosphate pathway. This NADPH pool serves as a critical cofactor for 11??-hydroxysteroid dehydrogenase type 1 (11??-HSD1), which catalyzes the reduction of inactive cortisone to active cortisol within the ER lumen. Thus, H6PD acts upstream of 11??-HSD1, and its activity is regulated by factors such as glucocorticoids, insulin, cAMP signaling, and oxidative stress. Downstream, H6PD-driven NADPH availability governs local cortisol production, which in turn modulates glucocorticoid receptor (GR) transactivation and the expression of glucocorticoid-responsive genes, including SGK1, GILZ, and FKBP5. Key interacting partners include 11??-HSD1 and the ER chaperones HSPA5 and CALR, which may contribute to proper enzyme folding or complex formation.
In T-47D cells, disruption of H6PD depletes ER luminal NADPH, thereby impairing 11??-HSD1-mediated cortisone-to-cortisol conversion. This reduction in local glucocorticoid activation blunts GR signaling and attenuates the transcription of target genes involved in cell proliferation, metabolism, and survival. Given the established crosstalk between glucocorticoid and estrogen receptor pathways in breast cancer, this H6PD knockout model provides a unique platform to investigate how altered cortisol synthesis impacts hormone-responsive tumor behavior, metabolic stress responses, and potential resistance mechanisms to endocrine therapies.
Key experimental applications include quantitative glucocorticoid metabolism analysis via LC-MS measurement of intracellular cortisol and cortisone, Western blot for 11??-HSD1 and GR, and RT-qPCR profiling of glucocorticoid-responsive transcripts such as SGK1, GILZ, and FKBP5. Functional assays such as GR transactivation reporter and proliferation assays (MTT or BrdU) evaluate phenotypic outcomes. The polyclonal population is also suitable for 11??-HSD1 inhibitor screening and exploring metabolic vulnerabilities in luminal A breast cancer. For further product details, pricing, or technical assistance, please contact Ascent Research.
