Cell death and cell-cycle arrest induced by incorporation of [ 3 H]thymidine into human haemopoietic cell lines

Abstract

Purpose : To investigate the influence of [ methyl - 3 H]thymidine ([ 3 H]Tdr) incorporated into human haemopoietic cell lines. Materials and methods : HL-60, Molt-4, Jurkat, Raji and SKW6-CL4 cells were incubated in the presence of [ 3 H]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 185kBq/ml [ 3 H]Tdr arrested at G 2 /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 [ 3 H]Tdr was incorporated into the human haemopoietic cell lines. Incorporation into HL-60 cells resulted in delay of S-phase progression, arrest at G 2 /M and apoptosis.