EFFECT OF Caspase-3 mRNA ANTISENSE OLIGODEOXYNUCLEOTIDES ON γ-IRRADIATION-INDUCED APOPTOSIS OF HL-60 CELLS

Objective To observe the inhibitory effect of caspase\|3 mRNA antisense oligodeoxynucleotides (ASODN) on the apoptosis of HL\|60 cells, and to screen the effective ASOND. Methods By means of the lipofectimine\|mediating transfection, caspase\|3 mRNA ASODNs of four different sequences were introduced into HL\|60 cells followed by γ irradiation.The gel electrophoresis was performed for the DNA ladder, Hoechst 33258\|propidium iodide fluorecent staining for the percentage of apoptotic cells, and FCM for quantitative analysis of apoptosis. Results When HL\|60 cells were transfected with caspase\|3 mRNA ASODNs targeting 5'noncoding region (sites -62 to -46) and initiative coding region (sites -1 to 16) at the final concentration of ≥3?μ mol/L, the DNA ladder was not detected with the gel electrophoresis, nor was the apoptotic peak found with FCM. Fluorescent staining analysis showed that the percentage of apoptotic cells was significantly lower than that in the control groups including non\|transfection group and mismatched oligodeoxynuleotide group ( P 0 01).With increase of the final transfection concentration, the inhibition rate of apoptosis was increased markedly. Furthermore, the noncoding region ASODN was more effective than the initiative coding region ASOND ( P 0 05).Conclusion Caspase\|3 mRNA ASODN can inhibit γ irradiation\|induced apoptosis of HL\|60 cells, the effect being sequence specific and dose dependent. [