The H6PD Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that disrupts the H6PD gene, encoding hexose-6-phosphate dehydrogenase, in the AGS human gastric adenocarcinoma cell line. This loss-of-function model provides a mixed population of edited cells ideal for pooled functional studies without requiring single-cell cloning.
AGS cells are adherent epithelial cells derived from a human gastric adenocarcinoma, widely used to investigate gastric cancer biology and Helicobacter pylori infection. They retain key features of gastric mucosal epithelial cells, including barrier and secretory functions, making them a relevant host for studying molecular pathways that regulate gastric epithelial homeostasis and transformation.
H6PD resides in the endoplasmic reticulum (ER) lumen, where it oxidizes glucose-6-phosphate to generate NADPH, the critical cofactor for luminal reductases such as 11??-hydroxysteroid dehydrogenase type 1 (HSD11B1). H6PD interacts with the glucose-6-phosphate transporter SLC37A4 and ER chaperones like HSPA5 (BiP). Its expression is regulated by the unfolded protein response factor XBP1, glucocorticoid receptor signaling, and cellular NADP+ levels. Downstream, H6PD-supplied NADPH drives HSD11B1-mediated conversion of cortisone to cortisol and supports ER protein disulfide isomerases and glutathione reductase, linking ER pentose phosphate pathway activity to glucocorticoid activation and redox homeostasis.
In AGS cells, H6PD knockout abolishes ER luminal NADPH generation, impairing HSD11B1-dependent cortisol reactivation and disturbing glucocorticoid signaling that can influence proliferation, survival, and stress responses. This model enables dissection of the ER-localized pentose phosphate pathway’s role in gastric cancer cell biology, including its contributions to ER stress mitigation and metabolic adaptation. It provides a tool to study how uncoupled ER redox control affects gastric mucosal functions and the interplay between glucocorticoid metabolism and oncogenic processes.
Typical applications include investigation of glucocorticoid metabolism in gastric epithelium, elucidation of ER redox regulation in cancer, and screening for 11??-HSD1 modulators. Assays such as western blotting, RT-qPCR, cortisol/cortisone LC-MS, NADPH measurement, 11??-HSD1 activity assays, and ER stress marker profiling can be employed. The cells also support metabolic disease modeling and pathway analysis for conditions like apparent cortisone reductase deficiency. Please contact Ascent Research for additional information or custom requests.
