Exposure of cells to ionizing radiation can cause apoptosis, Since antioxid
ants have been shown to protect against radiation-induced apoptosis, in thi
s study we have evaluated the putative protective effect of ascorbate again
st radiation-induced apoptosis as well as the production of peroxides in th
e cells. HL60 cells transport the oxidized form of ascorbic acid, dehydroas
corbic acid (DHA), and accumulate reduced ascorbate. Exposure of the cells
to 5-40 Gy X radiation resulted in induction of apoptosis, Preincubation of
the cells with DHA reduced the level of apoptosis after exposure to 5-20 G
y, Exposure of the cells to 5 or 20 Gy X radiation did not affect the intra
cellular concentration of peroxides, while phorbol myristate acetate (PMA),
which is known to induce production of H2O2 in cells (and served as a cont
rol), resulted in an increase in peroxides and a decrease in intracellular
ascorbate. Irradiation of the cells with 1-3 Gy resulted in up-regulation o
f expression of BCL2 without affecting the level of apoptosis, At higher do
ses of radiation, enhanced BCL2 expression did not prevent radiation-induce
d apoptosis, Loading of the cells with ascorbate prior to their exposure to
1-3 Gy X radiation did not affect the enhanced BCL2 expression observed in
the irradiated cells. At higher doses of radiation, ascorbate decreased ap
optosis and restored the level of BCL2 in the cells. Exposure of the cells
to 3-20 Gy X radiation enhanced the cell surface expression of TNFRSF6 (for
merly known as Fas/APO-1) antigen and enhanced anti-TNFRSF6 antibody-induce
d apoptosis of the cells. Ascorbate loading did not affect expression of TN
FRSF6 and did not overcome the anti-TNFRSF6 antibody-induced apoptosis, In
conclusion, our data demonstrate that exposure of HL60 cells to radiation e
nhanced BCL2 and TNFRSF6 expression. Ascorbate did not affect BCL2 or TNFRS
F6 expression. We therefore conclude that it protects HL60 cells against ra
diation-induced apoptosis, although the mechanisms of protection must still
be elucidated. (C) 1999 by Radiation Research Society.