Deoxyadenosine toxicity in an adenosine deaminase-inhibited human CCRF-CEM T-lymphoblastoid cell line causes cell swelling.
The human T-lymphoblastoid cell line CCRF-CEM, pre-treated with 2'-deoxycoformycin, was used as a model for adenosine deaminase deficiency to investigate how 2'-deoxyadenosine exerts its cytotoxic effects. Incubation of these cells with 1 microM or 5 microM deoxyadenosine for 24 and 48 h caused an increase of up to 50% in their modal cell volume as measured by a Coulter Size Distribution Analyzer and this increase in cell volume was accompanied by an increase in their fragility and deformability. The swelling of cells was concomitant with the phosphorylation of deoxyadenosine and its intracellular accumulation as dATP. There was no evidence of osmotic imbalance or of inhibition of the Na+/K(+)-dependent ATPase activity as the intracellular concentrations (and the intracellular:extracellular ratios) of Na+, K+ and Ca2+ were essentially unchanged. Cytochalasin B (20 microM) also caused lymphoblasts to swell over a 6-h period and its effect on cell size was similar to that of either 1 microM or 5 microM deoxyadenosine over 24 or 48 h. Longer time-courses of incubation with cytochalasin B caused severe toxicity leading to the death and lysis of a significant proportion of the cells. Other drugs, such as colchicine, vincristine and vinblastine that are known to affect various components of the cytoskeleton also caused swelling of cells in a concentration- and time-dependent manner but there was no evidence that these effects were additive or synergistic with those of deoxyadenosine. Inhibition of DNA synthesis, either directly by aphidicolin or indirectly by hydroxyurea, was less cytotoxic than the effect caused by deoxyadenosine. We conclude that one of the toxic effects resulting from the excessive phosphorylation of deoxyadenosine and its accumulation as dATP in human T-lymphoblasts is not dependent on inhibition of DNA synthesis but may be caused by the disruption of the cytoskeleton in these cells.
Bagnara
A S
AS
School of Biochemistry and Molecular Genetics, University of New South Wales, Kensington, Australia.
McDonald
L E
LE
Slade
H M
HM
D000243:Adenosine Deaminase / Q000172:deficiency*; D002118:Calcium / Q000032:analysis; D002460:Cell Line / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D003599:Cytoskeleton / Q000187:drug effects; D004261:DNA Replication / Q000187:drug effects; D003838:Deoxyadenine Nucleotides / Q000032:analysis*; D003839:Deoxyadenosines / Q000633:toxicity*; D006801:Humans; D011092:Polyethylene Glycols; D011188:Potassium / Q000032:analysis; D012964:Sodium / Q000032:analysis; D013601:T-Lymphocytes / Q000201:enzymology / Q000473:pathology*
D003838:Deoxyadenine Nucleotides; D003839:Deoxyadenosines; D011092:Polyethylene Glycols; D012964:Sodium; D000243:Adenosine Deaminase; D011188:Potassium; D002118:Calcium