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The Neurogenic Compound P7C3 Regulates the Aerobic Glycolysis by Targeting Phosphoglycerate Kinase 1 in Glioma
Wenjin Chen1,2, Weiqiang Jia3, Cuiying Wu2
1The Second School of Clinical Medicine, Southern Medical University, Guangzhou, China.
Background:
P7C3 is a neurogenic compound that exhibits neuroprotective properties in neural cells. However, its target proteins and effects in glioma are unknown.
Methods:
The candidate P7C3 target proteins were analyzed using a human protein microarray containing 23136 human proteins. A streptavidin agarose affinity assay was used to verify the direct interaction between P7C3 and phosphoglycerate kinase 1 (PGK1). Mass spectrometry was used to identify the binding sites of PGK1 for P7C3 binding. Seahorse XF96 extracellular flux analyzer was used to measure the cell oxygen consumption rate and extracellular acidification rate. Glycolytic metabolites were measured using the related kits. Protein level was detected by western blotting and immunohistochemical staining. Autophagy was analyzed using a transmission electron microscope and western blotting. The malignancy of tumor progression in vitro and in vivo was analyzed based on cell viability, apoptosis and proliferation, migration and invasion, and xenograft model. Glial cells were marked by antibodies via immunohistochemical staining.
Results:
The human protein microarray identified 577 candidate P7C3 target proteins. The global profile of P7C3 target proteins indicated that P7C3 regulates glycolysis. Metabolic experiments confirmed that P7C3 regulates aerobic glycolysis in glioma cells. The underlying mechanism of P7C3 was found to be direct targeting PGK1 at lysine residues and asparagine residues, and the specific P7C3-PGK1 interaction led to decreased protein level and total intracellular kinase activity of PGK1. The Cancer Genome Atlas and Chinese Glioma Genome Atlas databases indicated that the mRNA level of PGK1 is significantly increased in high-grade glioma, and the abnormally high mRNA level of PGK1 is associated with a poor prognosis in patients with glioma, suggesting that PGK1 is a promising target for glioma therapy. The inhibition of PGK1 and the subsequent suppression of aerobic glycolysis caused by P7C3 inhibited the malignant growth of glioma in vitro and in vivo. Furthermore, P7C3 did not damage normal glial cells under concentration, which exhibit an inhibitory effect on gliomas.
Conclusions:
This study revealed that P7C3 suppresses glioma by regulating aerobic glycolysis via directly targeting PGK1. Furthermore, we identified the P7C3 target proteins for the first time which is expected to provide scientific clues for future studies.
Insights
P7C3 compound suppresses glioma by targeting phosphoglycerate kinase 1 (PGK1), inhibiting aerobic glycolysis and tumor growth. This neurogenic compound shows promise for glioma therapy by selectively targeting cancer cells.
Area of Science:
- Neuroscience
- Oncology
- Biochemistry
Background:
- P7C3 is a known neurogenic and neuroprotective compound.
- Its specific protein targets and mechanisms in glioma remain uncharacterized.
Purpose of the Study:
- To identify P7C3 target proteins in glioma.
- To elucidate the molecular mechanisms by which P7C3 affects glioma progression.
- To evaluate the therapeutic potential of P7C3 in glioma treatment.
Main Methods:
- Protein microarray analysis to identify P7C3 binding partners.
- Biochemical assays (streptavidin agarose, mass spectrometry) to confirm direct interaction with PGK1.
- Metabolic assays (Seahorse analyzer) to measure glycolysis.
- Western blotting and immunohistochemistry for protein analysis.
- Cell viability, apoptosis, proliferation, migration, and invasion assays.
- In vivo xenograft models and transmission electron microscopy for autophagy analysis.
Main Results:
- Identified 577 candidate P7C3 target proteins, revealing P7C3's role in regulating glycolysis.
- Confirmed P7C3 directly targets phosphoglycerate kinase 1 (PGK1), reducing its protein levels and kinase activity.
- Demonstrated that elevated PGK1 mRNA levels correlate with poor prognosis in glioma patients.
- Showed P7C3 inhibits glioma cell growth, migration, and invasion in vitro and in vivo by suppressing aerobic glycolysis.
- P7C3 exhibited selective toxicity towards glioma cells without harming normal glial cells.
Conclusions:
- P7C3 suppresses glioma progression by directly targeting PGK1 and inhibiting aerobic glycolysis.
- PGK1 is a potential therapeutic target for glioma.
- This study provides the first identification of P7C3 target proteins, offering insights for future research.
