The H6PD Knockout 143B Polyclonal Cells product consists of a polyclonal population of 143B human osteosarcoma cells genetically modified by CRISPR/Cas9-mediated disruption of the H6PD gene (hexose-6-phosphate dehydrogenase). This product provides a heterogeneous loss-of-function model, enabling researchers to examine the consequences of H6PD ablation without clonal selection artifacts. The polyclonal format ensures a diverse representation of editing events across the population, suitable for studying pooled effects on cellular pathways.
The 143B cell line is a well-established model of human osteosarcoma, derived from a bone tumor and retaining features of malignant osteoblast-like cells. It is widely employed in cancer biology to investigate tumor proliferation, metastasis, and metabolic reprogramming. As a bone cancer model, 143B exhibits aggressive growth and is particularly valuable for dissecting the molecular interactions within the tumor microenvironment and evaluating therapeutic vulnerabilities.
H6PD resides in the endoplasmic reticulum (ER) and catalyzes the first step of the pentose phosphate pathway, oxidizing glucose-6-phosphate to generate NADPH. This cofactor is essential for the reductase activity of 11??-hydroxysteroid dehydrogenase type 1 (11??-HSD1), which converts inactive cortisone into active cortisol. Cortisol then binds to the glucocorticoid receptor (GR), triggering transcriptional regulation of metabolic target genes such as phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase). H6PD function is interconnected with the glucose-6-phosphate transporter (G6PT) and is modulated by upstream signals including TNF-?? and IL-1??, linking inflammatory cytokines to local glucocorticoid activation.
In the osteosarcoma context, disruption of H6PD is anticipated to alter NADPH availability and impair cortisol production, thereby perturbing GR-mediated gene expression programs that control proliferation, apoptosis, and migration. This model provides a unique platform to explore how metabolic shifts in the ER influence cancer cell behavior, potentially uncovering novel dependencies on glucocorticoid synthesis. Additionally, the knockout cells can serve as an in vitro model for cortisone reductase deficiency, a disorder characterized by impaired 11??-HSD1 activity, and may offer insights into metabolic syndrome and insulin resistance, as H6PD-driven NADPH generation is critical for hepatic gluconeogenesis.
Researchers can employ the H6PD Knockout 143B Polyclonal Cells in a broad range of experiments. Quantification of cortisol levels by ELISA and NADPH measurement can directly assess the impact on glucocorticoid metabolism. Cell proliferation and migration assays enable functional evaluation of H6PD in osteosarcoma aggressiveness. Transcriptomic profiling via RNA-seq, combined with western blotting or RT-qPCR for downstream targets like PEPCK and G6Pase, facilitates mechanistic dissection of GR signaling. Drug sensitivity studies can identify compounds that exploit the knockout-induced metabolic vulnerability. For further details on validation and availability, please contact Ascent Research.
