CAJ | 학술논문

目的观察小剂量地西他滨(DAC)单用或联合柔红霉素(DNR)对白血病细胞株HL-60的凋亡作用及对抑癌基因PTEN mRNA表达的影响.方法以不同浓度的DAC、DNR单药及联合处理HL-60细胞株,应用四甲基偶氮唑蓝(MTT)法、流式细胞术检测不同时相药物对细胞的凋亡作用;采用实时荧光定量聚合酶链反应(FQ-PCR)技术检测不同浓度组中HL-60细胞PTEN mRNA表达.结果 DAC、DNR单药对HL-60细胞的生长抑制作用呈剂量、时间依赖性,联合用药组的抑制作用则更为明显[72 h抑制率为(80.23±1.71)％,P＜ 0.001];5.0 μmol/L DAC联合1.0μmol/L DNR作用72 h细胞凋亡率最高,抑制作用均较DAC和DNR单药组明显(F=30.199,P＜ 0.001);不同浓度DAC与DNR联用后,细胞PTEN mRNA表达量增加,呈浓度依赖性,明显高于对照组及DNR单药组(F=578.218,P＜ 0.001).结论 DAC能显著抑制HL-60细胞的增殖并诱导其凋亡,与DNR联合对HL-60细胞增殖抑制作用有协同效应,其机制可能与DAC的去甲基化作用使PTEN mRNA的表达量增加有关.
목적 관찰소제량지서타빈(DAC)단용혹연합유홍매소(DNR)대백혈병세포주HL-60적조망작용급대억암기인PTEN mRNA표체적영향.방법 이불동농도적DAC、DNR단약급연합처리HL-60세포주,응용사갑기우담서람(MTT)법、류식세포술검측불동시상약물대세포적조망작용;채용실시형광정량취합매련반응(FQ-PCR)기술검측불동농도조중HL-60세포PTEN mRNA표체.결과 DAC、DNR단약대HL-60세포적생장억제작용정제량、시간의뢰성,연합용약조적억제작용칙경위명현[72 h억제솔위(80.23±1.71)％,P＜ 0.001];5.0 μmol/L DAC연합1.0μmol/L DNR작용72 h세포조망솔최고,억제작용균교DAC화DNR단약조명현(F=30.199,P＜ 0.001);불동농도DAC여DNR련용후,세포PTEN mRNA표체량증가,정농도의뢰성,명현고우대조조급DNR단약조(F=578.218,P＜ 0.001).결론 DAC능현저억제HL-60세포적증식병유도기조망,여DNR연합대HL-60세포증식억제작용유협동효응,기궤제가능여DAC적거갑기화작용사PTEN mRNA적표체량증가유관.
Objective To observe the cell proliferation inhibition of DNA methyltransferase (DNMT) inhibitors decitabine (DAC) combined with daunorubicin (DNR) in human leukemia cell line HL-60.Methods The effects of DNR and DAC were examined in HL-60 cells by cell viability using MTT method,and cell death using flow cytometric (FCM).Results DAC,DNR single drug application showed their effects on cell proliferation was dependent of dose and time,the inhibition effect of combined treatment group was much clearer [inhibition ratio of 72 hours was (80.23±1.71) ％,P ＜ 0.001].The highest apoptosis rate was at 5.0 μmol/L DAC combined with 1.0 μmol/L DNR for 72 hours,which was statistic significant (F =30.199,P ＜ 0.001).Combinations of different concentrations of DAC and DNR increased expression of PTEN mRNA in concentration-dependent manner,which was significantly higher than the control group and DNR single drug group (F =578.218,P ＜ 0.001).Conclusions DAC can significantly inhibit the proliferation of HL-60 cells and induce apoptosis,synergistic effect can be observed when DAC combined with DNR.The underlying mechanism can be due to DAC demethylation effect to increase PTEN mRNA expression.