羟基喜树碱诱导HL-60细胞凋亡的研究

目的 :探讨国产羟基喜树碱 (HCPT)对 HL - 6 0细胞的作用机制和规律。方法 :应用细胞形态学检查 ,DNA凝胶电泳及流式细胞仪分析检测。结果 :HCPT 1mg/ L和 5 mg/ L作为 HL - 6 0细胞 4h细胞凋亡率分别为 5 0 .6 %和 6 8.6 %。光镜检查见细胞核固缩、碎裂 ;电镜检查见染色质向核周边凝集。 DNA电泳显示典型的梯状条带。当 HCPT浓度超过 5 mg/ L或作用超过 4h,细胞凋亡率无明显增加 ,出现饱和现象。HL - 6 0细胞经 HCPT作用后 ,S期细胞显著减少 ,G0 / G1 期细胞增加。结论 :HCPT诱导 HL - 6 0细胞出现凋亡并具有饱和效应 ,HCPT为 S期特异性药物。临床上 HCPT应与细胞周期非特异性药物联合用药。

APOPTOSIS IN HL-60 CELLS INDUCED BY HYDROXYLCAMPTOTHECIN

Objective:To study the mechanism through which hydroxylcamptothecin (HCPT) made in china killed HL 60 cells and the law that HCPT killed the cells.Methods:By checking the morphology of apoptotic cells,agarose gel electrophoresis of DNA fragmentation and quantitation of apoptotic cells by means of flow cytometric analysis.Results:The percentage of apoptotic cells was 50.6% and 68.6% after the cells were incubated with 1 mg/L or 5 mg/L HCPT for 4 hours,respectively.Chromatin condensation and nuclear fragmentation were seen in the cells treated with HCPT by light and electron microscopy.DNA electrophoresis of HCPT treated cells showed nuclear fragmentation in oligonuclesosmal fragments(DNA ladder).The percentage of apoptotic cells neither increased obviously with growth in the concentration of HCPT more than 5mg/L,nor with prolongation in the duration of the cells incubated with HCPT exceeding 4 hours.After the cells were treated with HCPT,the number of the cells in S phase decreased greathly,nevertheless,the amount of the cells in G 0/G 1 phase increased.Conclusion:HCPT induces apoptosis in HL 60 cells and it has "saturation".HCPT is the drug killed the cells in S phase in HL 60 cells specially.The results suggest that HCPT should be applied with other drugs whose effects are non special on the phases in HL 60 cells.