Uncoupling of incorporation of ascorbic acid and apoptosis induction

Exposure of human promyelocytic leukemic HL-60 cells to millimolar concentration of sodium ascorbate induced apoptotic cell death. The extent of apoptosis induction was a positive function of temperature at the time of exposure. The incorporation of [1-14C] ascorbic acid into the cytosolic fraction of HL-60 cells was also temperature-dependent, and competitively inhibited by active analogs (L-ascorbic acid, sodium L-ascorbate, D-isoascorbic acid, sodium 6-beta-O-galactosyl-L-ascorbate, sodium 5,6-benzylidene-L-ascorbate), but not by inactive analogs (L-ascorbic acid-2-phosphate magnesium, L-ascorbic acid 2-sulfate). Calcium depletion, which had considerably reduced the apoptosis-inducing activity of sodium ascorbate, did not affect the intracellular incorporation of [14C] ascorbic acid. These data suggests that cell death might not be simply induced by the intracellular incorporation of ascorbate, but rather initiated by the rapid elevation of intracellular Ca2+ concentration, possibly mediated by an as yet unidentified temperature-sensitive mechanism.