The H6PD Knockout PaTu 8988t Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the human PaTu 8988t pancreatic ductal adenocarcinoma line. This product provides a mixed cell pool carrying H6PD gene disruptions, enabling study of endoplasmic reticulum glucose-6-phosphate dehydrogenase activity without clonal isolation. It is engineered to interrogate H6PD’s role in NADPH supply and glucocorticoid activation in a metastatic pancreatic cancer background.
PaTu 8988t cells were established from a liver metastasis of pancreatic ductal adenocarcinoma and serve as a model for aggressive, metastatic disease. They retain malignant epithelial features, including aberrant signaling and metabolic reprogramming. Using this host line provides a clinically relevant context to examine how H6PD loss influences pancreatic cancer cell biology, especially glucocorticoid metabolism and redox balance.
H6PD encodes a hexose-6-phosphate dehydrogenase that catalyzes the first step of the pentose phosphate pathway in the ER, converting glucose-6-phosphate to 6-phosphogluconolactone and generating NADPH. This NADPH is required by HSD11B1 (11??-hydroxysteroid dehydrogenase type 1) to convert cortisone to cortisol. H6PD activity is governed by the NADP+/NADPH ratio, glucose-6-phosphate levels, and insulin signaling, and it interacts with the glucose-6-phosphate transporter SLC37A4 and ER chaperones. Disruption of H6PD impairs NADPH delivery to HSD11B1, diminishing glucocorticoid activation and affecting NADPH-dependent reductases like glutathione reductase, thereby compromising oxidative stress defense.
In PaTu 8988t cells, H6PD knockout disrupts ER NADPH regeneration critical for HSD11B1 activity and local cortisol production. This likely alters redox homeostasis, sensitizing cells to oxidative damage and impairing glucocorticoid-dependent gene programs governing proliferation, migration, and drug resistance. The polyclonal knockout population is a valuable tool to dissect H6PD’s contribution to pancreatic tumor aggressiveness and to evaluate the H6PD?CHSD11B1 axis as a therapeutic target, alone or with chemotherapy.
Functional applications include 11??-HSD1 activity assays for cortisone-to-cortisol conversion, NADPH/NADP+ ratio measurement, and glucose-6-phosphate dehydrogenase activity assays. RT-qPCR and western blot confirm target disruption; 13C-glucose flux analysis traces pentose phosphate pathway activity; cortisol ELISA measures glucocorticoid output. Additional assays such as oxidative stress viability, migration/invasion screens, and drug sensitivity testing with HSD11B1 inhibitors are enabled. This polyclonal H6PD knockout in pancreatic cancer supports drug target validation, metabolic reprogramming studies, and glucocorticoid metabolism research. For further details, contact Ascent Research.
