Measurement of the ADP:ATP Ratio in Human Leukaemic Cell Lines Can Be Used as an Indicator of Cell Viability, Necrosis and Apoptosis

J Immunol Methods. 2000 Jun 23;240(1-2):79-92. doi: 10.1016/s0022-1759(00)00178-2.

Abstract

In this study the relative levels of ADP and ATP have been measured in cells undergoing apoptosis. Using HL60, CEM7, Jurkat and U937 cell lines and cytotoxic agents known to induce apoptosis, there was a significant correlation (P<0.01 for all models) between the ADP:ATP ratio and the degree of apoptosis measured by TUNEL and estimation of the sub G(0) fraction by propidium iodide staining and flow cytometry. The ratio measured in viable proliferating cells was found to be less than 0.11 compared with ratios between 0.11 and 1.0 seen in cells undergoing apoptosis. The higher the percentage of hypodiploidy the greater the ratio. Necrosis induced by heat shock resulted in ADP:ATP ratios in excess of 15.0. When primary cultures of AML blast cells were used, there was again a significant correlation between the ADP:ATP ratio and the degree of hypodiploidy. Recent evidence suggests that apoptosis is accompanied by opening of the mitochondrial permeability pores, leading to disruption of the mitochondrial transmembrane potential (DeltaPsi(m)). This results in caspase activation due to the release of cytochrome c and apoptogenic factors into the cytosol. In five experiments using CEM7 and dexamethasone the mitochondrial transmembrane potential was assessed using the fluorescent cyanine dye JC-1 and flow cytometry. Functioning mitochondria concentrate the JC-1 to produce red fluorescence. Loss of mitochondrial transmembrane potential results in green fluorescence only. The percentage of cells exhibiting red fluorescence correlated positively with the ATP values and negatively with the ADP:ATP ratio.

MeSH terms

  • Adenosine Diphosphate / analysis*
  • Adenosine Triphosphate / analysis*
  • Apoptosis
  • Camptothecin / pharmacology
  • Cell Death*
  • Cell Survival
  • Dexamethasone / pharmacology
  • Energy Metabolism*
  • HL-60 Cells
  • Humans
  • In Situ Nick-End Labeling
  • Jurkat Cells
  • Leukemia / metabolism*
  • Microscopy, Fluorescence
  • Necrosis
  • U937 Cells

Substances

  • Adenosine Diphosphate
  • Dexamethasone
  • Adenosine Triphosphate
  • Camptothecin