Cell death and cell-cycle arrest induced by incorporation of [3H]thymidine into human haemopoietic cell lines.
OBJECTIVE
To investigate the influence of [methyl-3H]thymidine ([3H]Tdr) incorporated into human haemopoietic cell lines.


METHODS
HL-60, Molt-4, Jurkat, Raji and SKW6-CL4 cells were incubated in the presence of [3H]Tdr. Cell proliferation, cell viability, DNA fragmentation and expression of caspase-3 and Bcl-2 families were examined. The cell-cycle of HL-60 was analysed using flow cytometry.


RESULTS
In HL-60, Molt-4 and Jurkat, cell death was accompanied by DNA nucleosomal fragmentation and activation of caspase-3. In Raji and SKW6-CL4, it was accompanied by neither. Protein levels of Bcl-2 and Bad in HL-60 and Molt-4 did not significantly change, and that of Bax was decreased after a 3-day incubation. HL-60 incubated in the presence of 74 or 185 kBq/ml [3H]Tdr arrested at G2/M phase, and then underwent apoptosis. In 7.4 kBq/ml, the cell-cycle progressed after the delay in S-phase.


CONCLUSIONS
Two different modes of cell death were observed when [3H]Tdr was incorporated into the human haemopoietic cell lines. Incorporation into HL-60 cells resulted in delay of S-phase progression, arrest at G2/M and apoptosis.
Yanokura
M
M
Medical Research Institute, Tokyo Medical and Dental University, Japan.
Takase
K
K
Yamamoto
K
K
Teraoka
H
H
D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002352:Carrier Proteins / Q000096:biosynthesis; D020170:Caspase 1 / Q000378:metabolism; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects / Q000528:radiation effects*; D016923:Cell Death / Q000187:drug effects / Q000528:radiation effects; D002455:Cell Division / Q000187:drug effects / Q000528:radiation effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D053938:DNA Fragmentation / Q000187:drug effects; D004307:Dose-Response Relationship, Radiation; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000528:radiation effects*; D006801:Humans; D009842:Oligopeptides / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D013936:Thymidine / Q000493:pharmacokinetics / Q000494:pharmacology*; D014316:Tritium; D051028:bcl-2-Associated X Protein; D051025:bcl-Associated Death Protein
C494819:BAD protein, human; C494826:BAX protein, human; C109198:CIB1 protein, human; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors; D009842:Oligopeptides; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; C099158:acetyl-aspartyl-glutamyl-valyl-aspartal; D051028:bcl-2-Associated X Protein; D051025:bcl-Associated Death Protein; D014316:Tritium; C096429:L 709049; D011065:Poly(ADP-ribose) Polymerases; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D020170:Caspase 1; D013936:Thymidine