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Investigation of the Glycolytic Oscillator in Cancer Cells

L Che Fru

L Che Fru*, M Kissick , University of Wisconsin, Madison, WI


SU-I-GPD-T-648 (Sunday, July 30, 2017) 3:00 PM - 6:00 PM Room: Exhibit Hall

Purpose: We have used three fundamentally different techniques to observed oscillatory dynamics in oxygen partial pressure in xenograft tumors (UM-SCC22B cell line) that experience acute hypoxia. We have observed periods ranging from minutes to tens of minutes. The superposition of oxygen distribution and oscillatory dynamics have revealed that these oscillations have the propensity to occur in areas of low to intermediate oxygen partial pressure. These areas also coincide with hypoglycemic environments. This has led us to posit that the glycolytic oscillator, which is favored by hypoglycemia, is a cause of the oscillatory dynamics observed in acute hypoxia. The purpose of this research is to investigate the glycolytic oscillator in cancer cells in-vitro.

Methods: Cells will be transfected with plasmid DNA encoding the biosensor, Peredox-mCherry via a mammalian expression vector pcDNA3.1. Peredox-mCherry constitutes of a green fluorescent protein, a circularly permuted T-Sapphire, linked to a bacterial NADH-binding protein called Rex. Peredox-mCherry reports cytosolic NADH-NAD+ ratio in microscopic imaging of intact live cells with appropriate excitation energies and filters. NADH-NAD+ ratio is an indicator of glucose metabolism by glycolysis. These would be carried out in different concentrations of glucose, and in the presence and absence of a competitive inhibitor of phosphofructokinase.

Results: The results would potentially show that UM-SCC22B cells exhibit glycolytic oscillations, and the conditions necessary for it to occur. If the periods of oscillations are similar to those observed in the in-vivo experiments, this would be one step closer to pointing to the glycolytic oscillator as a cause of oscillations in acute hypoxia.

Conclusion: Results from these experiments would be applied to an in-vivo setup. Should a proven allosteric inhibitor of phosphofructokinase eliminate oscillations in acute hypoxia in a tumor, then this would be strong evidence that the glycolytic oscillator is a significant factor in the oscillations in acute hypoxia.

Funding Support, Disclosures, and Conflict of Interest: This investigation was supported by The National Cancer Institute of the NIH under award numbers T32 CA009206 and R00 CA160639 (RK), and in part by the University of Wisconsin Carbone Cancer Center (UWCCC) under award number NIH/NCI P30 CA014520- UWCCC-Support. Vevo2100LAZR system utilized was acquired using S10-OD018505 grant from NIH.

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