| Objective: Aim to investigate the apoptosis initiation phase in human leukemia HL-60 cells induced by DADS, establish the apoptosis initiation model.Methods: After incubation of HL-60 cells with 3.6mg·L-1 DADS, the effects of DADS on HL-60 cell growth inhibition were estimated by growth curve, inverted microscope, light microscope, flow cytometry method was used to determine the induction of apoptosis and the expression of actived caspase-3, DNA agarose electrophoresis was used to determine the induction of apoptosis, and protein expression of caspase-3, Bcl-2 was tested by western-blotting. Results: Growth curve show the growth of HL-60 cells could not be inhibited at day 1,but remarkably reduced the growth rate at day2 to day6 (24.1%, 36.5%, 44.2%,52%,53.6%,P<0.05), After HL-60 cells were exposed to DADS, the typical apoptotic morphological changes were observed under the light microscope and inverted microscope. Flow cytometry analysis showed that the apoptotic rate of DADS treated cells at day 1 and day 2 was 3.1%, 4.3%, with no difference from control cells(3.0%), while from day 3 to day 5, the apoptotic rate was 8.5%, 15.2% , 27.4% respectively. Have great difference from control cells, the DADS-withdrawal group from day 2 to day 5 the apoptotic rate was 7.9%, 12.4%,16.5%,18.8% respectively, significantly increased than the control group and the 1 day with DADS withdrawl(P<0.05).the expression of actived caspase-3 from day 2 to day 5 was 6.3%,10.0%,10.4%,14.9%,17.3% respectively, was higher than control group and DADS treated 1 day(P<0.05), DNA agarose electrophoresis was showed the DADS treated group from day 4 and the... |
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