Tumor cell culture survival following glucose and glutamine deprivation at typical physiological concentrations

Objective: Most glucose (and glutamine)-deprivation studies of cancer cell cultures focus on total depletion, and are conducted over at least 24 h. It is difficult to extrapolate findings from such experiments to practical anti-glycolytic treatments, such as with insulin-inhibiting diets (with 10%-50% carbohydrate dietary restriction) or with isolated limb perfusion therapy (which usually lasts about 90 min). The aim of this study was to obtain experimental data on the effect of partial deprivation of D-glucose and L-glutamine (to typical physiological concentrations) during 0 to 6-h exposures of HeLa cells. Methods: HeLa cells were treated for 0 to 6 h with 6 mM D-glucose and 1 mM c-glutamine (normal in vivo conditions), 3 mM D-glucose and 0.5 mM L-glutamine (severe hypoglycemic conditions), and 0 mM D-glucose and 0 mM L-glutamine ("starvation"). Polarization-optical differential interference contrast and phase-contrast light microscopy were employed to investigate morphologic changes. Results: Reduction of glucose levels from 6 to 3 mM (and glutamine levels from 1 to 0.5 mM) brings about cancer cell survival of 73% after 2-h exposure and 63% after 4-h exposure. Reducing glucose levels from 6 to 0 mM (and glutamine levels from 1 to 0 mM) for 4 h resulted in 53% cell survival. Conclusion: These data reveal that glucose (and glutamine) deprivation to typical physiological concentrations result in significant cancer cell killing after as little as 2 h. This supports the possibility of combining anti-glycolytic treatment, such as a carbohydrate-restricted diet, with chemotherapeutics for enhanced cancer cell killing. (C) 2014 Elsevier Inc. All rights reserved.