CAJ | 학술논문

目的研究低剂量地西他滨(DAC)联合伊马替尼(IM)对K562细胞株的增殖抑制作用及对bcr-abl表达的影响.方法单药及两药联合后,通过四甲基偶氮唑蓝(MTT)法观察药物对K562细胞株的增殖抑制作用,流式细胞术检测药物对K562细胞株早期凋亡率及细胞周期,巢式反转录-聚合酶链反应(RT-PCR)半定量检测药物对K562细胞株bcr-abl mRNA表达.结果 DAC与IM单药对K562细胞的抑制作用呈浓度时间依赖性.两药联合用药抑制作用较单药组明显(F=43.947、165.580、321.193、296.101,均P<0.05),24、48、72 h各浓度组与对照组比较差异均有统计学意义(F=202.759、168.457、417.538,均P<0.05).DAC及IM单药作用药物对K562细胞株均使G,期细胞明显增多,IM0.2μmol/L作用于K562细胞株48 h可见6.7%早期凋亡细胞,IM 0.2 μmol/L联合DAC 4μmol/L早期凋亡细胞增加至8.4%.bcr-abl mRNA表达水平降低,DAC 4 μmol/L作用48 h后可降低K562细胞中bcr-abl mRNA表达(约14%),IM 0.2 μmol/L降低约40%,联合用药表达量明显降低(约60%).联合用药组与单药组比较差异有统计学意义(F=71.981,P<0.05).结论 DAC对K562细胞的增殖抑制作用与细胞周期阻滞、诱导凋亡及降低bcr-abl mRNA表达有关,两药联合可显著抑制K562细胞增殖.
목적 연구저제량지서타빈(DAC)연합이마체니(IM)대K562세포주적증식억제작용급대bcr-abl표체적영향.방법 단약급량약연합후,통과사갑기우담서람(MTT)법관찰약물대K562세포주적증식억제작용,류식세포술검측약물대K562세포주조기조망솔급세포주기,소식반전록-취합매련반응(RT-PCR)반정량검측약물대K562세포주bcr-abl mRNA표체.결과 DAC여IM단약대K562세포적억제작용정농도시간의뢰성.량약연합용약억제작용교단약조명현(F=43.947、165.580、321.193、296.101,균P<0.05),24、48、72 h각농도조여대조조비교차이균유통계학의의(F=202.759、168.457、417.538,균P<0.05).DAC급IM단약작용약물대K562세포주균사G,기세포명현증다,IM0.2μmol/L작용우K562세포주48 h가견6.7%조기조망세포,IM 0.2 μmol/L연합DAC 4μmol/L조기조망세포증가지8.4%.bcr-abl mRNA표체수평강저,DAC 4 μmol/L작용48 h후가강저K562세포중bcr-abl mRNA표체(약14%),IM 0.2 μmol/L강저약40%,연합용약표체량명현강저(약60%).연합용약조여단약조비교차이유통계학의의(F=71.981,P<0.05).결론 DAC대K562세포적증식억제작용여세포주기조체、유도조망급강저bcr-abl mRNA표체유관,량약연합가현저억제K562세포증식.
Objective Imatinib mesylate (IM) is the most active agent in treating chronic myeloid leukemia (CML). 5-Aza-2-deoxycytidine (DAC) is a cytosine analogue that inhibits DNA methylation and the activity in myeloid leukemia. Therefore,we investigated combining these two drugs in human leukemia cell line K562. Methods The effects of IM and DAC was examined in K562 cells including cell viability using MTT method,cell cycle phase and cell death using flow cytometric (FCM),and the expression of bcr-abl mRNA by RT-PCR. Results Both DAC and IM resulted in time and concentration-dependent induction of cell death. DAC and IM in combination produced a greater inhibition of growth against K562 cells (F =43.947,165.580,321.193,296.101,P<0.05). The main effect and interaction between two drugs was statistically significant (F = 202.759,168.457,417.538,P <0.001) after 24 h,48 h,72 h and a greater reduction in expression of bcr-abl mRNA than either agent alone. The difference was statistically significant (F =71.981,P <0.05). The number of G1 phase cells were increased significantly when induced by single agent. 48 h incubation with IM 0.2 μmol/L alone or combined with DAC 4 μmol/L showed 6.7 %,8.4 % pre-apoptosis cells,respectively. After incubation for 48 h with DAC 4 μmol/L,the expression of mRNA were decreased by 14 %,IM 0.2 μmol/L showed 40 % reduction,and combination group were significantly depressed for the mRNA expression by 60 %. Conclusion The combination of DAC and IM showed synergistic effects on cell death in K562 cells. These data suggested that DAC used in combination with IM has clinical potential in the treatment of chronic myeloid leukemia.